Team:BNDS China/Notebook

BNDS CHINA2019

Week1-Week2

We prepared our experiment materials at the beginning, then transform our plasmids into competent cells. We did colony PCR and plasmid PCR for sequencing and preparing enough pSB4C5, pSB1C3-lac, pSB1C3-tEF1, pSB1C3-tADH1, pYES, OKS, pYES/NTA, pSB1C3-lac+todA, todA, pSB1C3-lac, todB for the future experiments. Then, we used the todA, todB, todD and OKS to do bacteria transformation.

Week3

We constructed plasmids pSB1C3-lac-todC1C2, pSB1C3-lac-laccase, pYES-OKS-tTDH-pTEF-ZhuI, pYES-OKS-tTDH-pHHF-ZhuJ, then we did bacteria transformation with these plasmids. We also did colony PCR for sequencing pSB1C3-lac-todA, pSB1C3-lac-todB, pSB1C3-lac-todD and pYES-OKS.

Week4

We did colony PCR with pSB1C3-lac-todD, pSB1C3-lac-todC1C2 and plasmid PCR for pSB1C3-lac, which are all sent to company for sequencing. After that, we constructed pSB1C3-lac-todA, pSB1C3-lac-todB, pSB1C3-lac-todC1C2 and pSB1C3-lac-todD. Then we did bacteria transformation for pSB1C3-lac-todA, pSB1C3-lac-todB, pSB1C3-lac-todC1C2 and pSB1C3-lac-todD.

Week5-Week8

We constructed pYES-OKS-tTDH-pTEF-ZhuI-tENO-pHHF-ZhuJ, GFP+ and GFP- and did bacteria transformation with them. After that, we did yeast transformation with our pYES-OKS, pYES-OKS-tTDH-pTEF-ZhuI, pYES-OKS-tTDH-pTEF-ZhuI, pYES-OKS-tTDH-pTEF-ZhuI-tENO-pHHF-ZhuJ, GFP+ and GFP- plasmids. We also did protein purification for pSB1C3-lac-todA, pSB1C3-lac-todB, pSB1C3-lac-todC1C2 and pSB1C3-lac-todD.