Team:Beijing HDFLS High/Project

From 2014hs.igem.org

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=TITLE=
=TITLE=
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The UV-responsing generators for a wintergreen odor and protective melittin
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The protective melittin generator;
 +
The UV-responsing tandem generators for a wintergreen odor and protective melittin.
 +
 
=ABSTRACT=
=ABSTRACT=
-
We design a bacteria that produces a wintergreen odor and protective melittin under extensive UV radiation. The biological system includes  two generators: the UV-responsing wintergreen odor generator (BBa_K994000) and UV-responsing melittin generator. The melittin generator is composed of two transcriptional devices:  RecA (SOS) promoter (BBa_J22106) and synthesized melittin gene with enterokinase digested sequence. The protective generator takes as input extensive UV radiation and produces as output fusion melittin. Melittin  can improve immunity and scavenging free radicals to alleviate the harm from electromagnetic radiation, such as UV and blue light.
+
The protective melittin generator is composed of two transcriptional devices:  RecA (SOS) promoter (BBa_J22106) and synthesized melittin gene with enterokinase digested sequence. The protective generator takes as input extensive UV radiation and produces as output fusion melittin. Melittin  can improve immunity and scavenging free radicals to alleviate the harm from electromagnetic radiation, such as blue light and UV.
 +
[[File:The_protective_melittin_generator.png|center|400px]]
 +
We further construct a biological system that produces a wintergreen odor and protective melittin under extensive UV radiation. The biological system includes  two generators: the UV-responsing wintergreen odor generator (BBa_K994000) and  the protective melittin generator. It not only can warn people of intense UV irradiation by a wintergreen odor, but also produce protective melittin.
<br>
<br>
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[[File:Mellitin_generator.png|mid|600px]]
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[[File:Tandem Generators.png|center|800px]]
 +
 
 +
=BACKGROUND=
 +
===Blue light from computer induces eye retina producing free radical===
 +
 
 +
The electromagnetic spectrum, in order of increasing frequency and decreasing wavelength, can be divided, for practical engineering purposes, into radio waves, microwaves, infrared radiation, visible light, ultraviolet radiation, X-rays and gamma rays. The eyes of various organisms sense a relatively small range of frequencies of EMR called the visible spectrum or light. Higher frequencies (shorter wavelengths) correspond to proportionately more energy carried by each photon. Blue light is the most energetic portion of the visible light spectrum, it's less energetic than UV radiation but it also has the ability to penetrate into tissue and cause cellular damage.
 +
[[File:2000px-EM_spectrum.png|center|600px]]
 +
 
 +
Blue light is the hardest for the eye to focus on. We actually see a halo around bright blue light. Blue light scatters more inside the eyeball than other colors, producing more glare, more eyestrain and more fatigue. Intense blue light can cause damage to the retina, because blue is the hardest color for the retina to handle. The blue light induce that retinal produce free radicals. These radicals can lead to retinal pigment epithelial cell death. Then some light-sensitive cells lacks of nutrient, which results in visual impairment.
 +
[[File:Blue light.png|center|600px]]
 +
===Melittin has funcition of scavenging free radical===
 +
The old paint and song implies that the early ancient civilization was conscious of the possible virtues from bee stings.
 +
[[File:bee3.png|center|700px]]
 +
Melittin is the main component (50~55%) of bee venom,  which has multiple functions.
 +
[[File:Melittin3.png|center|600px]]
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=BACKGROUD=
 
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As the students in an international school, we always face many different kinds of assessment events and each of them need to finished by computer. After using Internet whole day, our eyes would feel uncomfortable with innumerable distinct kind radiations. How can we deal with it?
 
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The eye is a complex organ, composed of many different structures with varying sensitivity to the effects of radiation. An easy way to look at the eye is to divide it into zones that can be examined with respect to their susceptibility to radiation effects. It is also noteworthy that the effects of radiation on the eye not only depend on the sensitivity of the tissue itself but also on the mode of radiation delivery.
 
<br>
<br>
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[[File:Bees3.png|mid|600px]]
 
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==Step==
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[[File:Melittin function.png|center|600px]]
 +
 
 +
=PARTS=
 +
=I: Parts of Protective Melittin Generators.=
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[[image:part.png|center|400px]]
 +
 
 +
==Part A: Rec A (SOS) promoter==
 +
Extensive UV irradiation impings on heavily DNA damage and results in a single-strand DNA occuring, which activates RecA protein.Then,RecA protein leads to the expression of about 15 different repair proteins to rescue DNA damage.RecA protein is the central regulator in SOS repair system.Rec A (SOS) Promoter is available in registry part plate.
 +
[[Image: Part A.jpg|center|400px]]
 +
 
 +
==Part B: Fusion melittin ==
 +
We synthesized fusion melittin gene with enterokinase digestion site. Melittin has the function of scavenging free radicals.
 +
[[Image:melittin.jpg|center|700px]]
 +
 
 +
=II: Parts of Tandem Generators.=
 +
==Part A: Wintergreen Odor Generator==
 +
The generator was constructed last year, and  can produce wintergreen odor under extensive UV radiation
 +
 
 +
==Part B: Protective Melittin Genertor==
 +
The generator is constructed this year, and can produce protective melittin under extensive UV radiation
 +
 
 +
=METHODS=
 +
===METHOD I: The Protective Melittin Generator===
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[[Image:method I.jpg|center|800px]]
 +
 
 +
===METHOD II: The Tandem Generators===
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We construct tandem Generators, using similar method with protective melittin generator
 +
 
 +
=RESULTS=
 +
===Result I:The Protective Melittin Generator===
 +
=====Plates=====
 +
[[Image:Result Plate 2.jpg|center|800px]]
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=====Electrophoresis Map after Restriction Digest=====
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 +
[[Image:Result Map4.jpg|center|900px]]
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=====DNA Sequencing and Submitting Part=====
 +
http://parts.igem.org/Part:BBa_K1283000
 +
[[Image:1283000.jpg|center|800px]]
 +
 
 +
===Result II:Tandem Generators===
 +
=====Plates=====
 +
[[Image:Result IIPlate .jpg|center|400px]]
 +
 
 +
=====Electrophoresis Map after Restriction Digest=====
 +
 
 +
[[Image:Result electrophoresis .jpg|center|600px]]
 +
 
 +
===== DNA Sequencing and Submitting Part=====
 +
http://parts.igem.org/Part:BBa_K1283002
 +
[[Image:1283002.jpg|center|800px]]
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===Step1===
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=STEPS=
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We have to got Part A and Part B's DNA before we digestion.
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===Step1 Transformation and Miniprep===
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Part A will be used after transformation and miniprep.
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We get Part A plasmid from registry, and construct Part B plasmid.
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We got Part B by artificial synthesis.
+
[[File:step1.png|center|600px]]
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===Step2===
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===Step2 Digestion===
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Digestion
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[[File:Step2.png|center|600px]]
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===Step3===
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===Step3 Ligation===
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Electrophoresis
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[[File:step3.png|center|600px]]
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===Step4===
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===Step4 Tansformation, Miniprep and Digestion again===
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Ligation
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[[File:step4.png|center|600px]]
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===Step5===
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===Step5 Sequencing===
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Sequencing
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Latest revision as of 12:45, 20 June 2014

Header wiki.png

Beijing HDFLS iGEM 2013

 



Contents

TITLE

The protective melittin generator; The UV-responsing tandem generators for a wintergreen odor and protective melittin.

ABSTRACT

The protective melittin generator is composed of two transcriptional devices: RecA (SOS) promoter (BBa_J22106) and synthesized melittin gene with enterokinase digested sequence. The protective generator takes as input extensive UV radiation and produces as output fusion melittin. Melittin can improve immunity and scavenging free radicals to alleviate the harm from electromagnetic radiation, such as blue light and UV.

The protective melittin generator.png

We further construct a biological system that produces a wintergreen odor and protective melittin under extensive UV radiation. The biological system includes two generators: the UV-responsing wintergreen odor generator (BBa_K994000) and the protective melittin generator. It not only can warn people of intense UV irradiation by a wintergreen odor, but also produce protective melittin.

Tandem Generators.png

BACKGROUND

Blue light from computer induces eye retina producing free radical

The electromagnetic spectrum, in order of increasing frequency and decreasing wavelength, can be divided, for practical engineering purposes, into radio waves, microwaves, infrared radiation, visible light, ultraviolet radiation, X-rays and gamma rays. The eyes of various organisms sense a relatively small range of frequencies of EMR called the visible spectrum or light. Higher frequencies (shorter wavelengths) correspond to proportionately more energy carried by each photon. Blue light is the most energetic portion of the visible light spectrum, it's less energetic than UV radiation but it also has the ability to penetrate into tissue and cause cellular damage.

2000px-EM spectrum.png

Blue light is the hardest for the eye to focus on. We actually see a halo around bright blue light. Blue light scatters more inside the eyeball than other colors, producing more glare, more eyestrain and more fatigue. Intense blue light can cause damage to the retina, because blue is the hardest color for the retina to handle. The blue light induce that retinal produce free radicals. These radicals can lead to retinal pigment epithelial cell death. Then some light-sensitive cells lacks of nutrient, which results in visual impairment.

Blue light.png

Melittin has funcition of scavenging free radical

The old paint and song implies that the early ancient civilization was conscious of the possible virtues from bee stings.

Bee3.png

Melittin is the main component (50~55%) of bee venom, which has multiple functions.

Melittin3.png


Melittin function.png

PARTS

I: Parts of Protective Melittin Generators.

Part.png

Part A: Rec A (SOS) promoter

Extensive UV irradiation impings on heavily DNA damage and results in a single-strand DNA occuring, which activates RecA protein.Then,RecA protein leads to the expression of about 15 different repair proteins to rescue DNA damage.RecA protein is the central regulator in SOS repair system.Rec A (SOS) Promoter is available in registry part plate.

Part A.jpg

Part B: Fusion melittin

We synthesized fusion melittin gene with enterokinase digestion site. Melittin has the function of scavenging free radicals.

Melittin.jpg

II: Parts of Tandem Generators.

Part A: Wintergreen Odor Generator

The generator was constructed last year, and can produce wintergreen odor under extensive UV radiation

Part B: Protective Melittin Genertor

The generator is constructed this year, and can produce protective melittin under extensive UV radiation

METHODS

METHOD I: The Protective Melittin Generator

Method I.jpg

METHOD II: The Tandem Generators

We construct tandem Generators, using similar method with protective melittin generator

RESULTS

Result I:The Protective Melittin Generator

Plates
Result Plate 2.jpg
Electrophoresis Map after Restriction Digest
Result Map4.jpg
DNA Sequencing and Submitting Part

http://parts.igem.org/Part:BBa_K1283000

1283000.jpg

Result II:Tandem Generators

Plates
Result IIPlate .jpg
Electrophoresis Map after Restriction Digest
Result electrophoresis .jpg
DNA Sequencing and Submitting Part

http://parts.igem.org/Part:BBa_K1283002

1283002.jpg

STEPS

Step1 Transformation and Miniprep

We get Part A plasmid from registry, and construct Part B plasmid.

Step1.png

Step2 Digestion

Step2.png

Step3 Ligation

Step3.png

Step4 Tansformation, Miniprep and Digestion again

Step4.png

Step5 Sequencing

Retrieved from "http://2014hs.igem.org/Team:Beijing_HDFLS_High/Project"