Difference between revisions of "Team:Fudan-TSI/Composite Part"

For best composite part, we would like to present you with our Cre recombinase together with degradation tag. Cre recombinase (BBa_K3257044) is responsible for the recombination process between the reverse transcribed cDNA and the target Gene of Interest (GOI), which is rather one of the most important processes in our system. Because of the leakage of uninduced Cre expression, we have spent much effort in lowering the steady-state expression level of Cre. An important approach is to attach 5 different types of degradation tags after the CDS of Cre. Using EGFP (BBa_E0040) as a reporter, we can see that the degradation rate of degradation tag with amino acid residual sequence of AANDENWVLAA (BBa_K3257073) is moderate for Cre recombinase to reach a steady but relatively low level of expression. Transformed into BL21 (DE3) with another plasmid containing lox sites and analyzed by colony PCR, we can see that homologous recombination happens between the same lox sites instead of gene knockout (Figure 1). For more information of other composite parts, please refer to our collections and the iGEM Part Registry ranging from BBa_K3257100 to BBa_K3257118.