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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:26.0pt;
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><img width=258 height=194
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| - | src="https://static.igem.org/mediawiki/2014hs/e/e4/Image0001.jpg" align=left hspace=12><u><span
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| - | lang=EN-US style='font-size:16.0pt;color:black'>1) Reporter Systems Overview</span></u><u><span
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| - | lang=EN-US style='font-size:11.5pt;color:black'>:</span></u></p>
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| - |
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| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
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| - | margin-left:0cm;text-align:justify;line-height:normal'><span lang=EN-US
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| - | style='font-size:11.5pt;color:black; '>a) Fluorescent Proteins:
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| - | The green fluorescent protein (GFP) is a </span><a
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| - | href="http://en.wikipedia.org/wiki/Protein"><span lang=EN-US style='font-size:
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| - | 11.5pt;color:black; ;text-decoration:none'>protein</span></a><span
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| - | lang=EN-US style='font-size:11.5pt;color:black; '> that
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| - | exhibits bright green </span><a href="http://en.wikipedia.org/wiki/Fluorescence"><span
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| - | lang=EN-US style='font-size:11.5pt;color:black; ;text-decoration:
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| - | none'>fluorescence</span></a><span lang=EN-US style='font-size:11.5pt;
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| - | color:black; '> when exposed to light in the blue to </span><a
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| - | href="http://en.wikipedia.org/wiki/Ultraviolet"><span lang=EN-US
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| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>ultraviolet</span></a><span
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| - | lang=EN-US style='font-size:11.5pt;color:black; '> range.
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| - | Although many other marine organisms have similar green fluorescent proteins,
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| - | GFP traditionally refers to the protein first isolated from the </span><a
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| - | href="http://en.wikipedia.org/wiki/Jellyfish"><span lang=EN-US
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| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>jellyfish</span></a><span
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| - | lang=EN-US style='font-size:11.5pt;color:black; '> </span><a
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| - | href="http://en.wikipedia.org/wiki/Aequorea_victoria"><span lang=EN-US
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| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>Aequorea
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| - | victoria</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
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| - | '>. The GFP from A. victoria has a major </span><a
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| - | href="http://en.wikipedia.org/wiki/Fluorescence_spectroscopy"><span lang=EN-US
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| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>excitation
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| - | peak</span></a><span lang=EN-US style='font-size:11.5pt;color:black; '> at a </span><a href="http://en.wikipedia.org/wiki/Wavelength"><span
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| - | lang=EN-US style='font-size:11.5pt;color:black; ;text-decoration:
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| - | none'>wavelength</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
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| - | '> of 395 nm and a minor one at 475 nm. Its emission peak is at
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| - | 509 nm, which is in the lower green portion of the </span><a
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| - | href="http://en.wikipedia.org/wiki/Visible_spectrum"><span lang=EN-US
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| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>visible
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| - | spectrum</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
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| - | '>. The fluorescence </span><a
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| - | href="http://en.wikipedia.org/wiki/Quantum_yield"><span lang=EN-US
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| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>quantum
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| - | yield</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '> (QY) of GFP is 0.79. The GFP from </span><img width=143
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| - | height=143 src="https://static.igem.org/mediawiki/2014hs/3/3b/Image0002.jpg" align=right hspace=12><span
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| - | lang=EN-US style='font-size:11.5pt;color:black; '>the </span><a
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| - | href="http://en.wikipedia.org/wiki/Sea_pansy"><span lang=EN-US
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| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>sea
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| - | pansy</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
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| - | '> (</span><span lang=EN-US style='font-size:12.0pt'><a
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| - | href="http://en.wikipedia.org/wiki/Renilla_reniformis"><span style='font-size:
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| - | 11.5pt;color:black; ;text-decoration:none'>Renilla reniformis</span></a></span><span
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| - | lang=EN-US style='font-size:11.5pt;color:black; '>) has a single
| |
| - | major excitation peak at 498 nm. </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black; '>In </span><a
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| - | href="http://en.wikipedia.org/wiki/Cell_biology"><span lang=EN-US
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| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>cell</span></a><span
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| - | lang=EN-US style='font-size:11.5pt;color:black; '> and </span><a
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| - | href="http://en.wikipedia.org/wiki/Molecular_biology"><span lang=EN-US
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| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>molecular
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| - | biology</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '>, the GFP </span><a href="http://en.wikipedia.org/wiki/Gene"><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; ;text-decoration:
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| - | none'>gene</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '> is frequently used as a </span><a
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| - | href="http://en.wikipedia.org/wiki/Reporter_gene"><span lang=EN-US
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| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>reporter
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| - | of expression</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '>. In modified forms it has been used to make </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Biosensor"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>biosensors</span></a><span
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| - | lang=EN-US style='font-size:11.5pt;color:black; '>, and many
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| - | animals have been created that express GFP as a proof-of-concept that a gene
| |
| - | can be expressed throughout a given organism. The GFP gene can be introduced
| |
| - | into organisms and maintained in their genome through breeding, injection with
| |
| - | a </span><a href="http://en.wikipedia.org/wiki/Viral_vector"><span lang=EN-US
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| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>viral
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| - | vector</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '>, or cell transformation. To date, the GFP gene has been
| |
| - | introduced and expressed in many </span><a
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| - | href="http://en.wikipedia.org/wiki/Bacteria"><span lang=EN-US style='font-size:
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| - | 11.5pt;color:black; ;text-decoration:none'>Bacteria</span></a><span
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| - | lang=EN-US style='font-size:11.5pt;color:black; '>, </span><a
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| - | href="http://en.wikipedia.org/wiki/Yeast"><span lang=EN-US style='font-size:
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| - | 11.5pt;color:black; ;text-decoration:none'>Yeast</span></a><span
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| - | lang=EN-US style='font-size:11.5pt;color:black; '> and other </span><a
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| - | href="http://en.wikipedia.org/wiki/Fungi"><span lang=EN-US style='font-size:
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| - | 11.5pt;color:black; ;text-decoration:none'>Fungi</span></a><span
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| - | lang=EN-US style='font-size:11.5pt;color:black; '>, fish (such
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| - | as </span><a href="http://en.wikipedia.org/wiki/Zebrafish"><span lang=EN-US
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| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>zebrafish</span></a><span
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| - | lang=EN-US style='font-size:11.5pt;color:black; '>), plant, fly,
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| - | and mammalian cells, including human. </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Martin_Chalfie"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>Martin
| |
| - | Chalfie</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '>, </span><a
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| - | href="http://en.wikipedia.org/wiki/Osamu_Shimomura"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>Osamu
| |
| - | Shimomura</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '>, and </span><a
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| - | href="http://en.wikipedia.org/wiki/Roger_Y._Tsien"><span lang=EN-US
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| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>Roger
| |
| - | Y. Tsien</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
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| - | '> were awarded the 2008 </span><a
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| - | href="http://en.wikipedia.org/wiki/Nobel_Prize_in_Chemistry"><span lang=EN-US
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| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>Nobel
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| - | Prize in Chemistry</span></a><span lang=EN-US style='font-size:11.5pt;
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| - | color:black; '> on 10 October 2008 for their discovery and
| |
| - | development of the green fluorescent protein. </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
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| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
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| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoListParagraph style='margin-bottom:0cm;margin-bottom:.0001pt;
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| - | text-align:justify;text-indent:-18.0pt;line-height:normal'><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black'>a)<span style='font:7.0pt "Times New Roman"'>
| |
| - | </span></span><span lang=EN-US style='font-size:11.5pt;color:black'> Luciferin:<span
| |
| - | style=' '> Luciferin (from the Latin </span>lucifer,
| |
| - | "light-bringer") is a generic term for the light-emitting </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Chemical_compound"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>compound</span></a><span
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| - | lang=EN-US style='font-size:11.5pt;color:black; '> found in
| |
| - | organisms that generate </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Bioluminescence"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>bioluminescence</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '>. Luciferins
| |
| - | typically undergo an </span><img width=329 height=219
| |
| - | src="https://static.igem.org/mediawiki/2014hs/1/1e/Image0003.jpg" align=right hspace=12><a
| |
| - | href="http://en.wikipedia.org/wiki/Enzyme"><span lang=EN-US style='font-size:
| |
| - | 11.5pt;color:black; ;text-decoration:none'>enzyme</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '>-catalysed </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Oxidation"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>oxidation</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '> and the
| |
| - | resulting</span><a href="http://en.wikipedia.org/wiki/Excited_state"><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; ;text-decoration:
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| - | none'>excited state</span></a><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black; '> intermediate emits light upon decaying to its </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Ground_state"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>ground
| |
| - | state</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '>. This may refer to molecules that are substrates for both </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Luciferase"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>luciferases</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '> and </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Photoprotein"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>photoproteins</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '>. Luciferins
| |
| - | are a class of small-molecule </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Substrate_(biochemistry)"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>substrates</span></a><span
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| - | lang=EN-US style='font-size:11.5pt;color:black; '> that are </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Redox"><span lang=EN-US style='font-size:
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| - | 11.5pt;color:black; ;text-decoration:none'>oxidized</span></a><span
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| - | lang=EN-US style='font-size:11.5pt;color:black; '> in the
| |
| - | presence of the enzyme </span><a href="http://en.wikipedia.org/wiki/Luciferase"><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; ;text-decoration:
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| - | none'>luciferase</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '> to produce oxyluciferin and </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Energy"><span lang=EN-US style='font-size:
| |
| - | 11.5pt;color:black; ;text-decoration:none'>energy</span></a><span
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| - | lang=EN-US style='font-size:11.5pt;color:black; '> in the form
| |
| - | of </span><a href="http://en.wikipedia.org/wiki/Light"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>light</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '>. It is not
| |
| - | known just how many types of luciferins there are, but some of the
| |
| - | better-studied compounds are listed below. There are many types of luciferins,
| |
| - | yet all share the use of </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Reactive_oxygen_species"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>reactive
| |
| - | oxygen species</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '> to emit light. </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'>b) Xgal Reporter System:<span
| |
| - | style=' '> X-gal </span>is an </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Organic_compound"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>organic
| |
| - | compound</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '> consisting of</span><a
| |
| - | href="http://en.wikipedia.org/wiki/Galactose"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>galactose</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '> linked to a
| |
| - | substituted </span><a href="http://en.wikipedia.org/wiki/Indole"><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; ;text-decoration:
| |
| - | none'>indole</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '>. The compound was synthesized by </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Jerome_Horwitz"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>Jerome
| |
| - | Horwitz</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '> and collaborators in Detroit, MI, in 1964. The formal
| |
| - | chemical name is often shortened to less accurate but also less cumbersome
| |
| - | phrases such as bromochloroindoxyl galactoside. The X from </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Indoxyl"><span lang=EN-US style='font-size:
| |
| - | 11.5pt;color:black; ;text-decoration:none'>indoxyl</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '> may well be
| |
| - | the source of the X in the X-gal contraction. X-gal is much used in </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Molecular_biology"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>molecular
| |
| - | biology</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '> to test for the presence of an enzyme, </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Beta-galactosidase"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>β-galactosidase</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '>. It is also
| |
| - | used to detect activity of this enzyme in </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Histochemistry"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>histochemistry</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '> and </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Bacteriology"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>bacteriology</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '>. X-gal is one
| |
| - | of many indoxyl glycosides and esters that yield insoluble blue compounds
| |
| - | similar to </span><a href="http://en.wikipedia.org/wiki/Indigo"><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; ;text-decoration:
| |
| - | none'>indigo</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '> as a result of enzyme-catalyzed hydrolysis.</span><a
| |
| - | href="http://en.wikipedia.org/wiki/X-gal#cite_note-2"><sup><span lang=EN-US
| |
| - | style='font-size:7.0pt;color:black; ;text-decoration:none'>[2]</span></sup></a><sup><span
| |
| - | lang=EN-US style='font-size:7.0pt;color:black; '> </span></sup><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '>X-gal is an </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Structural_analog"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>analog</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '> of </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Lactose"><span lang=EN-US style='font-size:
| |
| - | 11.5pt;color:black; ;text-decoration:none'>lactose</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '>, and
| |
| - | therefore may be hydrolyzed by the </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Beta-galactosidase"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>β-</span></a><img
| |
| - | width=218 height=182 src="https://static.igem.org/mediawiki/2014hs/e/e8/Image0004.jpg" align=left
| |
| - | hspace=12><a href="http://en.wikipedia.org/wiki/Beta-galactosidase"><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; ;text-decoration:
| |
| - | none'>galactosidase</span></a><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black; '> enzyme which cleaves the β-</span><span
| |
| - | lang=EN-US style='font-size:12.0pt'><a
| |
| - | href="http://en.wikipedia.org/wiki/Glycosidic_bond"><span style='font-size:
| |
| - | 11.5pt;color:black; ;text-decoration:none'>glycosidic bond</span></a></span><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '> in D-lactose.
| |
| - | X-gal, when cleaved by β-galactosidase, yields galactose and
| |
| - | 5-bromo-4-chloro-3-hydroxyindole. The latter then spontaneously dimerizes and
| |
| - | is oxidized into 5,5'-dibromo-4,4'-dichloro-indigo, an intensely blue product
| |
| - | which is insoluble. X-gal itself is colorless, the presence of blue-colored
| |
| - | product therefore may be used as a test for the presence of an active
| |
| - | β-galactosidase. This easy identification of an active enzyme allows the
| |
| - | gene for β-galactosidase (the </span><span lang=EN-US style='font-size:
| |
| - | 11.5pt;color:black'>lacZ gene) to be used as a </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Reporter_gene"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>reporter
| |
| - | gene</span></a><span lang=EN-US style='font-size:11.5pt;color:black; '> in various applications. </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'>c) GUS Reporter System: <span
| |
| - | style=' '>The </span>GUS reporter system (GUS: </span><a
| |
| - | href="http://en.wikipedia.org/wiki/%CE%92-glucuronidase"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>β-glucuronidase</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '>) is a </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Reporter_gene"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>reporter
| |
| - | gene</span></a><span lang=EN-US style='font-size:11.5pt;color:black; '> system, particularly useful in plant </span><img width=167 height=249
| |
| - | src="https://static.igem.org/mediawiki/2014hs/5/51/Image0005.jpg" align=right hspace=12><a
| |
| - | href="http://en.wikipedia.org/wiki/Molecular_biology"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>molecular
| |
| - | biology</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '> and microbiology. Several kinds of GUS </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Reporter_gene_assay"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>reporter
| |
| - | gene assay</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '> are available, depending on the substrate used. The term </span><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black'>GUS staining refers to the most
| |
| - | common of these, a histochemical technique. The purpose of this technique is to
| |
| - | analyze the activity of a </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Promoter_(biology)"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>promoter</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '> (in terms of </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Gene_expression"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>expression</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '> of a gene
| |
| - | under that promoter) either in a quantitative way or through visualization of
| |
| - | its activity in different </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Biological_tissue"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>tissues</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '>. The
| |
| - | technique is based on </span><a
| |
| - | href="http://en.wikipedia.org/wiki/%CE%92-glucuronidase"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>β-glucuronidase</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '>, an enzyme
| |
| - | from the bacterium </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Escherichia_coli"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>Escherichia
| |
| - | coli</span></a><span lang=EN-US style='font-size:11.5pt;color:black; '>; this enzyme, when incubated with some specific colorless or non-fluorescent
| |
| - | </span><a href="http://en.wikipedia.org/wiki/Substrate_(biology)"><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; ;text-decoration:
| |
| - | none'>substrates</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '>, can transform them into </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Color"><span lang=EN-US style='font-size:
| |
| - | 11.5pt;color:black; ;text-decoration:none'>coloured</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '> or </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Fluorescence"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>fluorescent</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '> products. </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - | margin-left:0cm;text-align:justify;line-height:normal'><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; '> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-top:6.0pt;margin-right:0cm;margin-bottom:6.0pt;
| |
| - | margin-left:0cm;text-align:justify;line-height:normal'><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; '>d) Chloramphenicol
| |
| - | acetyltransferase (CAT) System: Chloramphenicol acetyltransferase (or CAT) is a
| |
| - | </span><a href="http://en.wikipedia.org/wiki/Bacteria"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>bacterial</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '> </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Enzyme"><span lang=EN-US style='font-size:
| |
| - | 11.5pt;color:black; ;text-decoration:none'>enzyme</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '> (</span><a
| |
| - | href="http://en.wikipedia.org/wiki/Enzyme_Commission_number"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>EC</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '> </span><a
| |
| - | href="http://enzyme.expasy.org/EC/2.3.1.28"><span lang=EN-US style='font-size:
| |
| - | 11.5pt;color:black; ;text-decoration:none'>2.3.1.28</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '>) that
| |
| - | detoxifies the </span><a href="http://en.wikipedia.org/wiki/Antibiotic"><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; ;text-decoration:
| |
| - | none'>antibiotic</span></a><a
| |
| - | href="http://en.wikipedia.org/wiki/Chloramphenicol"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>chloramphenicol</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '> and is
| |
| - | responsible for chloramphenicol resistance in bacteria. This enzyme covalently
| |
| - | attaches an</span><a href="http://en.wikipedia.org/wiki/Acetyl"><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; ;text-decoration:
| |
| - | none'>acetyl</span></a><span lang=EN-US style='font-size:11.5pt;color:black;
| |
| - | '> group from </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Acetyl-CoA"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>acetyl-CoA</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '> to
| |
| - | chloramphenicol, which prevents chloramphenicol from binding to </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Ribosome"><span lang=EN-US style='font-size:
| |
| - | 11.5pt;color:black; ;text-decoration:none'>ribosomes</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '>. A histidine
| |
| - | residue, located in the C-terminal section of the enzyme, plays a central role
| |
| - | in its catalytic mechanism.</span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black; '>The crystal structure of the type III enzyme from
| |
| - | </span><a href="http://en.wikipedia.org/wiki/Escherichia_coli"><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; ;text-decoration:
| |
| - | none'>Escherichia coli</span></a><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black; '> with chloramphenicol bound has been determined.
| |
| - | CAT is a trimer of identical subunits (monomer Mr 25,000) and the trimeric
| |
| - | structure is stabilised by a number of hydrogen bonds, some of which result in
| |
| - | the extension of a beta-sheet across the subunit interface. Chloramphenicol
| |
| - | binds in a deep pocket located at the boundary between adjacent subunits of the
| |
| - | trimer, such that the majority of residues forming the binding pocket belong to
| |
| - | one subunit while the catalytically essential histidine belongs to the adjacent
| |
| - | subunit. His195 is appropriately positioned to act as a general base catalyst
| |
| - | in the reaction, and the required tautomeric stabilisation is provided by an
| |
| - | unusual interaction with a main-chain carbonyl oxygen. CAT is used as a
| |
| - | reporter system to measure the level of a promoter or its tissue-specific
| |
| - | expression. The CAT assay involves monitoring acetylation of radioactively
| |
| - | labeled chloramphenicol on a </span><a
| |
| - | href="http://en.wikipedia.org/wiki/Thin_layer_chromatography"><span lang=EN-US
| |
| - | style='font-size:11.5pt;color:black; ;text-decoration:none'>TLC</span></a><span
| |
| - | lang=EN-US style='font-size:11.5pt;color:black; '> plate; CAT
| |
| - | activity is determined by looking for the acetylated forms of chloramphenicol,
| |
| - | which have a significantly increased migration rate as compared to the
| |
| - | unacetylated form.</span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'>d) Other Response Systems:
| |
| - | Some fast response systems were for constructed based on traditional response
| |
| - | systems like GFP or luciferin reporter system through protein engineering.
| |
| - | Although these response systems give fast response, they have high costs and/or
| |
| - | difficulty in implementation. The systems are devised by major genetics
| |
| - | companies that synthesize these systems using high cost equipment thus
| |
| - | reflecting the costs to the buyers resulting in very expensive prices making it
| |
| - | almost impossible for most of the iGEM teams to obtain them.</span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><u><span lang=EN-US style='font-size:16.0pt;
| |
| - | color:black'>2) Advantages of DegradEColor: </span></u></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'>a) <i>Our system aims to operate faster because it focuses on the
| |
| - | prencipal which is making the produced enzyme effect more substrata in which
| |
| - | are dyes instead of continuously producing new proteins. </i></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><i><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'>Our system works in a way that is suitable for different dyes and
| |
| - | peroxidase enzymes of the project it is going to be applied. </span></i></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'>c) <i>Also our system ables scientists to; develop dyes suitable
| |
| - | for their works, and chose the suitable enzyme for their subject living
| |
| - | material </i></span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><u><span lang=EN-US style='font-size:16.0pt;
| |
| - | color:black'>3) Disadvantages of DegradEColor: </span></u></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'>a) </span><span style='color:black'>Our protein is produced by
| |
| - | HRP-C enzyme which is taken by horseradish plant, and the other one LiP-1 is
| |
| - | taken by white rot fungus. Since our enzymes are taken from living material in
| |
| - | nature it might make things harder for people who wishes to use them on
| |
| - | different livings.</span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'>b) </span><span style='color:black'>The dyes we use alongside of
| |
| - | our enzymes origins are used in histologic and genetic studies in order to
| |
| - | colour DNA, RNA and other parts of cell, this situation is the reason why
| |
| - | our dyes does not to include the molecules we need which will increase
| |
| - | the efficiency of our project.</span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:16.0pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p style='margin:0cm;margin-bottom:.0001pt;text-align:justify'><u><span
| |
| - | lang=EN-US style='font-size:16.0pt;font-family:"Calibri","sans-serif";
| |
| - | color:black'>4) Planned Improvements of DegradEColor:</span></u></p>
| |
| - |
| |
| - | <p style='margin:0cm;margin-bottom:.0001pt;text-align:justify'><span
| |
| - | lang=EN-US style='font-size:16.0pt;font-family:"Calibri","sans-serif";
| |
| - | color:black'> </span><span lang=EN-US style='font-family:"Calibri","sans-serif";
| |
| - | color:black'>The dyes and enzymes we used should be engineered to be suitable
| |
| - | for our project if it is going to be used as a reliable reporter system which
| |
| - | will be used in synthetic biology and genetics in future. </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><u><span lang=EN-US style='font-size:16.0pt;
| |
| - | color:black'>a) Dye Improvements:</span></u></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:200%'><span lang=EN-US style='font-size:11.5pt;line-height:
| |
| - | 200%;color:black'>One of the actions that we conducted to further develop our
| |
| - | future plans was to meet up with organic chemistry experts specialized in the
| |
| - | field of dye synthesis. As a result of the meetings, we learned that it would
| |
| - | be possible to further develop the structure of our selected dyes in respect to
| |
| - | the speed of degradation thus reducing the total time of the process. </span><span
| |
| - | lang=EN-US style='font-size:12.0pt;line-height:200%;color:black'>. Our stops
| |
| - | were Prof. Dr. Cihangir Tanyeli from METU, and Doc. Dr. Zeynel Seferoğlu from
| |
| - | Gazi University. We took their thoughts about the possibility and any point
| |
| - | that can be advanced of our project. With this way, our project has been
| |
| - | approved and developed by experts.</span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><u><span lang=EN-US style='font-size:16.0pt;
| |
| - | color:black'>Prof. Dr. Cihangir Tanyeli</span></u></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><u><span lang=EN-US style='font-size:16.0pt;
| |
| - | color:black'>METU Faculty of Organic Chemistry</span></u></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><img width=389 height=292
| |
| - | src="https://static.igem.org/mediawiki/2014hs/9/98/Image0006.jpg" align=left hspace=12><span
| |
| - | lang=EN-US style='font-size:12.0pt;color:black'>One of the first questions we
| |
| - | asked to Mr Tanyeli was: What different technique or techniques can we increase
| |
| - | the effectiveness of Horse radish peroxidase and lignin peroxidase or how can
| |
| - | we make our dyes more sensitive and suitable through our enzymes and be able to
| |
| - | improve the usage field of our project?</span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'>He focused on re-designing the dyes to make them suitable for
| |
| - | enzymes which was his main field of study and he offered us 2 different
| |
| - | methods.</span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'>1st Method: We can develop the dye matter by using 4-amino-TEMPO.
| |
| - | This is a good redox system. By this way that we won’t need hydrogen peroxidase
| |
| - | for Oxidation</span><sub><span lang=EN-US style='font-size:7.5pt;color:black'> </span></sub><span
| |
| - | lang=EN-US style='font-size:12.0pt;color:black'>system. A polymer can be taken
| |
| - | as a Matrix. 4-amino-TEMPO can be immobilized to biodegradable polymer. With
| |
| - | the way that, sodium hypochlorite (NaClO) can be used as an alternative to
| |
| - | hydrogen peroxide. Since this is a macro molecular system, it can be
| |
| - | precipitated to water and regained this will provide us an ease of researching.
| |
| - | When used in a bulk system in the nature, most usable metalloporphirins have
| |
| - | silica like systems that has Si main structure, are more solid and hard to
| |
| - | degrade, but still, if the biodegradable systems are used catalytic activity
| |
| - | can be decreased and be a system that can fit to nature instead of being waste.</span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'>In order to block the degrading ability of the 4-amino-TEMPO
| |
| - | system, antioxidants like ascorbic acid can be used this happens by radical
| |
| - | group gets silenced by ascorbic acid which goes into a reaction with Bleach. He
| |
| - | also stated that we can use other antioxidants that alternatives of ascorbic
| |
| - | acid. In the other hand,. NaClO(Bleach instrument) is could be harmful for the
| |
| - | environment. 4 amino tempo system we suggested has an incredible balance with
| |
| - | bleach insturemuer so we for the nature we can chose the most envorment insrtie
| |
| - | , destroying its harms for environment because bleach + tempo system In
| |
| - | this method we focus on 4 amino tempos.. The reason we use amino instead of
| |
| - | hydroxide, we want our TEMPO molecule (oxidation instruments) can easily
| |
| - | interact with some organic molecules that can be produce by organisms with
| |
| - | amino group..</span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'>2</span><sup><span lang=EN-US style='font-size:7.5pt;color:black'>nd</span></sup><span
| |
| - | lang=EN-US style='font-size:12.0pt;color:black'> method: Some functional groups
| |
| - | can be placed on Carbon Nanotubes. Carboxylic acid can be placed on Carbon
| |
| - | Nanotubes with desiccation. Also 4-amino-TEMPO added on Carboxylic acid
| |
| - | functional group with direct covalent immobilization approach. If the 4-amino-TEMPO
| |
| - | stated gets inserted on and bleach instruments like NaClO added the environment
| |
| - | dye degradation will be happen. Big advantages of Carbon nanotubes that can be
| |
| - | regained with filtration and precipitation and this make this system more efficient
| |
| - | and economic and using over and over.</span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'>Both of this methods are using with switch systems that working
| |
| - | with pH or temperature control by an enzymes, or a bacteria or a bio systems.</span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'>We stated that time used by the systems used currently used around
| |
| - | the world takes 16 hours is decreased to a few hours by our system while we
| |
| - | were talking to our precious scholar. He gave us valuable information. We thank
| |
| - | to him for giving us time.</span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:13.0pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><u><span lang=EN-US style='font-size:16.0pt;
| |
| - | color:black'>Doc. Dr. Zeynel SEFEROĞLU</span></u></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><u><span lang=EN-US style='font-size:16.0pt;
| |
| - | color:black'>Gazi University Faculty of Organic Chemistry</span></u></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><img width=319 height=240
| |
| - | src="https://static.igem.org/mediawiki/2014hs/5/5c/Image0007.jpg" align=left hspace=12><span
| |
| - | lang=EN-US style='font-size:12.0pt;color:black'>We started our conservation
| |
| - | with our scholar how we can make our dyes more suitable for our dyes and make
| |
| - | it fit right in to our project. He suggested us to make a research on quantum
| |
| - | efficient dyes and increase our knowledge on functional fluorescence dyes. When
| |
| - | we first mentioned our project he liked it and stated that this could be used
| |
| - | easily and efficiently in genetic and synthetic biology world. He gave us some
| |
| - | articles that might help. He also stated that it is important to introduce to
| |
| - | science to the world since iGEM is already doing that we made someone else like
| |
| - | iGEM. We thank our scholar for giving us opportunity to have this conservation.</span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><u><span lang=EN-US style='font-size:16.0pt;
| |
| - | color:black; '>ALFA Dye</span></u><u><span lang=EN-US
| |
| - | style='font-size:16.0pt;color:black; '> and Chemistry</span></u></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><img width=238 height=394
| |
| - | src="https://static.igem.org/mediawiki/2014hs/f/f6/Image0008.jpg" align=left hspace=12><span
| |
| - | lang=EN-US style='font-size:12.0pt;color:black; '>We wanted to
| |
| - | inform the local companies about iGEM and our project. Our first target was a
| |
| - | professional corporation Alfa Dye. Here we talked about the possibility and
| |
| - | usage in daily life and received some information on industrial dyes. We
| |
| - | learned the dyes used in histologic dying are an addition to pigments as
| |
| - | connecting molecules. So if we want to add other activities other than
| |
| - | bioremediation of dye wastes we should add some small systems which will cleave
| |
| - | such connections. With this visit we had new a perspective through the future.
| |
| - | The company got happy for us to be in such competition. They sacrificed their
| |
| - | time in order to give us directions we thank them for that.</span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
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| - | color:black'> </span></p>
| |
| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
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| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><u><span lang=EN-US style='font-size:16.0pt;
| |
| - | color:black'>b) Enzyme and Genetic Dyes Improvements:</span></u></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
| |
| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:11.5pt;
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| - | color:black'>Another activity to further develop our future plans was to
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| - | consult genetic, biology and biochemistry experts and genetics companies to
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| - | assist in the development of the theory of our project. The feedback that we
| |
| - | received was indicating that through the development of the binding sites of
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| - | the enzymes with protein engineering the project could be improved and that the
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| - | project beared great potential for the improvision of synthetic biology.</span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:12.0pt;text-align:justify;line-height:
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| - | normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><u><span lang=EN-US style='font-size:16.0pt;
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| - | color:black'>Prof. Dr. Sadık Çiğdem</span></u></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><u><span lang=EN-US style='font-size:16.0pt;
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| - | color:black'>Tsukuba University Genetic Faculty Japan</span></u></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
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| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><img width=343 height=258
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| - | src="https://static.igem.org/mediawiki/2014hs/4/4e/Image0009.jpg" align=left hspace=12><span
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| - | lang=EN-US style='font-size:12.0pt;color:black'>He is one of the guest
| |
| - | scientist of Turgut Ozal University Genetic Department. When we mentioned him
| |
| - | about our project DegradEColor he is really impressed about the idea behind our
| |
| - | project and also he gave us some advice and information about our HRP enzyme.
| |
| - | He informed us that one of our enzyme HRP is used by ELISA and Western-Blot
| |
| - | experiments. HRP is added on antibodies in Western Blot and also ELISA and
| |
| - | cause the substrate ECL give response. We thanked to him for this opportunity. </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
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| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
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| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
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| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
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| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
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| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
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| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'> </span></p>
| |
| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><u><span lang=EN-US style='font-size:16.0pt;
| |
| - | color:black'>Prof. Dr. Esra Gündüz</span></u></p>
| |
| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><u><span lang=EN-US style='font-size:16.0pt;
| |
| - | color:black'>Turgut Özal University Faculty of Medicine</span></u></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'> </span></p>
| |
| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><img width=387 height=290
| |
| - | src="https://static.igem.org/mediawiki/2014hs/5/5d/Image0010.jpg" align=left hspace=12><span
| |
| - | lang=EN-US style='font-size:12.0pt;color:black'>We informed her about our iGEM
| |
| - | HS Project DegradEColor, she really interested on our project. She was very
| |
| - | pleased when she saw us, doing such work at this young age. She stated that she
| |
| - | will provide any support she can. </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
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| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
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| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
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| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
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| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
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| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
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| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
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| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'> </span></p>
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| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'> </span></p>
| |
| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'> </span></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'> </span></p>
| |
| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><u><span lang=EN-US style='font-size:16.0pt;
| |
| - | color:black'>Burak Yılmaz</span></u></p>
| |
| - |
| |
| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><u><span lang=EN-US style='font-size:16.0pt;
| |
| - | color:black'>The founder of SENTEGEN and its CEO</span></u></p>
| |
| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt;
| |
| - | color:black'> </span></p>
| |
| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
| |
| - | justify;line-height:normal'><img width=354 height=265
| |
| - | src="https://static.igem.org/mediawiki/2014hs/a/a7/Image0011.jpg" align=left hspace=12><span
| |
| - | lang=EN-US style='font-size:12.0pt;color:black'>We impressed Mr. Yılmaz with
| |
| - | our project and aim to reduce the time of response systems. He stated managing
| |
| - | time is a very big problem for companies. The company which will use our system
| |
| - | will manage their time 14~15 times more efficient. He said “This advantage can’t
| |
| - | be ignored”. He congratulated us for coming up with such a wonderful idea we
| |
| - | thank him for this.</span></p>
| |
| - |
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| - | <p class=MsoNormal style='margin-bottom:0cm;margin-bottom:.0001pt;text-align:
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| - | justify;line-height:normal'><span lang=EN-US style='font-size:12.0pt'> </span></p>
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| - |
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| - | <p class=MsoNormal style='text-align:justify'><span lang=EN-US
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| - | style='font-size:12.0pt;line-height:115%'><br>
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| - | <br>
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| - | <br>
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| - | <br>
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| - | </span></p>
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| - |
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| - | <p class=MsoNormal style='text-align:justify'><span style='font-size:13.0pt;
| |
| - | line-height:115%'> </span></p>
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| - |
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| - | </div>
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| - |
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| - | </body>
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| - | </head>
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| - | </html>
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