Team:JiangnanU China/Improve

Improvement

This year, we improved the sequence of the RBS BBa_B0033, and got the RBS BBa_K3137001 .

As we all know, RBS refers to a sputum-rich untranslated region upstream of the initiation codon AUG. BBa_B0033 is a weaker RBS based on Ron Weiss thesis. Compared with BBa_B0030, BBa_B0031, BBa_B0032, BBa_B0034, B0033 is weakest, and whose RBS strength is 0.35% of B0034. By analyzing their sequence, we found that B0033 has fewer purines and no not have AAA sequence. SO we designed the RBS by adjusting the proportion of bases.

Then we ligated the two RBS with promotor PrsmH (BBa_K3137000) and gene gfp (BBa_K3137011 ) and transform into E. coli BL21 to compare their green fluorescence brightness. The figure below shows that BBa_K3137001 has the stronger fluorescently protein expression than BBa_B0033, that means it’s efficient to increase the protein expression by increasing the number of purines in the RBS sequence, which is beneficial to the binding of ribosomes.