Why Promoter and Terminator design?

Our project tests the limits to which biology can be standardized. Towards this goal, the endogenous promoter of every essential gene will be replaced with one of seven standard promoters producing different expression levels. The promoters are biobrick parts and are currently being functionally tested. To determine which promoter to pair with which gene, microarray expression data from the literature was used. The terminator of every essential gene will also be replaced with a standard biobrick terminator.

Advantages of standardized promoters include:

• streamlined amplifaction of genes from genomic DNA, as amplicons can all start precisely at the start codon
< potential for easy manipulation of gene expression levels: the same gene part can easily be assembled with different promoters to test the effect of different expression levels
• no concerns about which transcription factors need to be retained in the essential gene list to ensure essential genes are transcibed
• as the location and properties of many enhancers are unknown, using all standardize promoters results in a much better characterized, well understood system, which is more amenabel to alteration