Team:Gifu/Improve
Improvement plan
*Caution: We did not submit the part to iGEM HQ this year. Since linear DNA is workable, we evaluate how much important the enhancer is for PURE system and T7 RNA polymerase. We will submit this part in the competition of 2019. Thus this part is not met the gold criteria.
As proof of improvement of previous composite part of iGEM, BBa_K1491033 was provided by iGEM HQ. Typical alignment of T7 promoter is used in this part. However, in PURE system, the enhancer of the stem-loop structure need to be located in the downstream of T7 promoter to make an expression of protein intense. Thus we also design the part (Seq.2) below to show the improvement and this may contribute to the development of synthetic biology and iGEM competition. In Fig.1 we showed the detail of T7 promoter we used this year. Translation enhancer upstream of the SD sequence of mRNA, enhance the biosynthesis of protein. The enhancer may contribute to a direct interaction with ribosome protein S1. Fig.1 shows the detail of the enhancer.
*Caution: BBa_K1332002 contains histidine tag. This time we excluded histidine tag. The function of the coding of this sequence is exactly the same as BBa_E1010. In Seq.2 it has s stop codon.
Improvement Result
As a result of the experiment, we can see the clear fluorescence in our new part, but not in BBa_K1491033.
In addition, we can see the apparent difference in SDS-PAGE.
