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Team:BostonU/FusionProteins
From 2014.igem.org
Need for Fusion ProteinsFusion 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. Making Fusion ProteinsFusion 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. Testing Fusion ProteinsFusion 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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