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Team:Acton-BoxboroughRHS/Research
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E.coli has a higher pH range and may be easier to assemble, but may be less productive. Yeast will probably be more productive in the long run but is otherwise inferior
we need an open reading frame (orf) a ribosome binding site (rbs), promoter, and terminator at least 3-5 enzymes: pepsin, salivary amylase, trypsin, chymotripsin, pancreatic amylase
This image shows the template for a typical plasmid requirement
A promoter is a DNA sequence that tends to recruit transcriptional machinery and lead to transcription of the downstream DNA sequence.
A ribosome binding site (RBS) is an RNA sequence found in mRNA to which ribosomes can bind and initiate translation.
.Protein domains are portions of proteins cloned in frame with other proteins domains to make up a protein coding sequence. Some protein domains might change the protein's location, alter its degradation rate, target the protein for cleavage, or enable it to be readily purified.
The Terminator is the 'last' region of the plasmid, where RNA polymerase stops transcription. There are two main types of terminators in prokaryotes, rho-dependent terminators, and rho-independent terminators. Rho-dependent termination bases its function on the Rho protein, which causes the RNA polymerase to fall disassociate form the plasmid. Rho-independent termination does not use Rho to disassociate the RNA polymerase, but instead stops by the DNA sequence, here's how it works. The DNA sequence contains a two regions that are rich in cytosine and guanine. When this is transcribed, those regions on the RNA form hydrogen bonds to itself: effectively forming a 'loop' which disassociates the DNA polymerase from the DNA.
A plasmid is a circular, double-stranded DNA molecules typically containing a few thousand base pairs that replicate within the cell independently of the chromosomal DNA. A plasmid backbone is defined as the plasmid sequence beginning with the BioBrick suffix, including the replication origin and antibiotic resistance marker, and ending with the BioBrick prefix.
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