Difference between revisions of "Part:BBa K2542014"
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<partinfo>BBa_K2542014 short</partinfo> | <partinfo>BBa_K2542014 short</partinfo> | ||
| − | 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([https://parts.igem.org/Part:BBa_K775004 BBa_K775004]) of the previous team to make the expression efficiency | + | 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([https://parts.igem.org/Part:BBa_K775004 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 ]）. |
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
<partinfo>BBa_K2542014 SequenceAndFeatures</partinfo> | <partinfo>BBa_K2542014 SequenceAndFeatures</partinfo> | ||
| + | ===Experiment and Result=== | ||
| + | <p> | ||
| + | iGEM18_NEFU_China creat a new parts [https://parts.igem.org/Part:BBa_K2542014 BBa_K2542014]，which is based on [https://parts.igem.org/Part:BBa_K775004 BBa_K775004]. [https://parts.igem.org/Part:BBa_K2542014 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 [https://parts.igem.org/Part:BBa_K775004 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. | ||
| + | </p> | ||
| + | |||
| + | [[Image:T--NEFU_China--Kozak_sequence_change_result.jpg]] | ||
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Latest revision as of 07:59, 14 October 2018
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.
