Part:BBa_K2765005

promoter of gene GSH1

This part consist of the promoter GSH1 (GSH1p), which is an inducible promoter (the promoter of glutamate--cysteine ligase) from Saccharomyces cerevisiae. In this project, we use it as the downstream gene promoter of Yap1 to turn on the expression of dCas9.We have 9 different promoters as option.

We obtained the promoter through PCR. Here is the electropherogram that we gained the promoters.

Because we have no idea of the exact strength of these promoters, we chose both strong promoters and weak ones in this result and compared them with promoters that we have already known their strength. We tested downstream genes of Yap1 whose names were: CTT1, GLR1, TRX2, TRR1, TSA1, SOD2, GSH1, and GSH2, together with strong promoter gal1p and weak promoter msy1p.

We obtained these promoters through PCR, and we ligated these promoters with gfp gene to express GFP protein and test their strength through fluorescence intensity. This is the first step to screen the proper promoter. The results of the fluorescence intensity are as follows.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 1012
  Illegal XhoI site found at 1904
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]