Part:BBa_K2542014
pFUS1-EGFP-tCYC
This plasmid allows a type yeast to sense the alpha factor secreted by alpha-type yeast and express green fluorescent protein. We modified the part(BBa_K775004) of the previous team to make the expression efficiency weaker，and make it more appropriate for suicide gene（[http://2018.igem.org/Team:NEFU_China/Suicide Information Destroyed ]）.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Experiment and Result
iGEM18_NEFU_China creat a new parts BBa_K2542014，which is based on BBa_K775004. BBa_K2542014 is more suitable for somewhere which needs low expression in yeast.It's allows a type yeast to sense the α factor secreted by α-type yeast and express green fluorescent protein. We modified the part BBa_K775004 of the previous team by turning mutating the sequence “AAAAAA” into “GCCACC” to make the expression efficiency weaker，and make it more appropriate for suicide gene（[http://2018.igem.org/Team:NEFU_China/Suicide Information Destroyed ]). We constructed two plasmids with different kozak sequences and transferred them into yeast. Then, we use SD-Trp medium to culture yeast, and add with the solution which containing α factor. The solution containing α factor is obtain by extracting the liquid supernatant of the culture medium for culturing α-type yeast. After we add 200ul the α factor sloution to each one and culture yeast for 20 hours，we measured the expression of GFP in yeast by flow cytometry.
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