Difference between revisions of "Part:BBa K2542014"

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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE 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.

@@ Line 3: / Line 3: @@
 <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 weaker，and make it more appropriate for suicide gene（[http://2018.igem.org/Team:NEFU_China/Suicide Suicide]）.
+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 ]）.
 <!-- Add more about the biology of this part here
@@ Line 11: / Line 11: @@
 <span class='h3bb'>Sequence and Features</span>
 <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]]
 <!-- Uncomment this to enable Functional Parameter display