Team:BostonU/FusionProteins

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Revision as of 21:25, 23 July 2014

Fusion Proteins

Need for Fusion Proteins (Bigger Font) Fusion Proteins are fused coding sequences that allow us to use one transcriptional unit (with a repressor and reporter protein fused together) rather than two units (one with the repressor and the other with the reporter protein, whose expression depends on the expresser of the repressor). Fusion Proteins play a very important part in Project Chimera. In addition to helping us reduce the total transcriptional units used, they also allow for greater expression of tandem coding sequences. If we look at constructs that have repeating sequences of ribosome binding sites and reporter proteins in one transcriptional unit, a peculiar effect can be observed. When ribosomes bind to the genes sequentially, the ribosome that binds before might block the next ribosome from binding to the mRNA and translating the gene. So, the later genes are expressed less than the genes before. This problem is solved with fused proteins as only one ribosome will then be required to translate the entire sequence, eliminating any possible problems during translation.

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-             <th scope="col"><br>Fusion proteins are analogous to tandem promoters. For our purposes, we fused multiple repressors with reporter proteins in order to check whether a transcriptional unit containing fusion proteins worked just as well as two transcriptional units (one with the repressor and the other with the reporter protein). These constructs were fused by adding unique fusion sites to their ends and then, ligating them using the <a href='https://2014.igem.org/Team:BostonU/MoClo'>Modular Cloning</a> method. The objective of making this library was to help us create standard transcriptional units that could universally be used to make intricate genetic circuits.</th> </tr>
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+Need for Fusion Proteins (Bigger Font)
+Fusion Proteins are fused coding sequences that allow us to use one transcriptional unit (with a repressor and reporter protein fused together) rather than two units (one with the repressor and the other with the reporter protein, whose expression depends on the expresser of the repressor).
+Fusion Proteins play a very important part in Project Chimera. In addition to helping us reduce the total transcriptional units used, they also allow for greater expression of tandem coding sequences.
+If we look at constructs that have repeating sequences of ribosome binding sites and reporter proteins in one transcriptional unit, a peculiar effect can be observed. When ribosomes bind to the genes sequentially, the ribosome that binds before might block the next ribosome from binding to the mRNA and translating the gene. So, the later genes are expressed less than the genes before. This problem is solved with fused proteins as only one ribosome will then be required to translate the entire sequence, eliminating any possible problems during translation.</th> </tr>