Difference between revisions of "Part:BBa K2765005"
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<partinfo>BBa_K2765005 short</partinfo> | <partinfo>BBa_K2765005 short</partinfo> | ||
| − | This part consist of the promoter | + | This part consist of the promoter GSH1 (GSH1p), which is an inducible promoter (the promoter of glutamate--cysteine ligase) from <i>Saccharomyces cerevisiae</i>. 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. | We obtained the promoter through PCR. Here is the electropherogram that we gained the promoters. | ||
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[[Image: T--BIT-China--iGEM2018-PartFeedback-realgel.png |center|400px|]] | [[Image: T--BIT-China--iGEM2018-PartFeedback-realgel.png |center|400px|]] | ||
| − | 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: | + | 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. | + | We obtained these promoters through PCR, and we ligated these promoters with <i>gfp</i> 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. |
[[Image: T--BIT-China--iGEM2018-PartsFeedback-experimentresult.png |center|400px|]] | [[Image: T--BIT-China--iGEM2018-PartsFeedback-experimentresult.png |center|400px|]] | ||
Latest revision as of 21:14, 17 October 2018
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1012
Illegal XhoI site found at 1904 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]