Difference between revisions of "Part:BBa K2922039"
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<table><tr><th>[[Image:chromoP.png|thumb|180px|Fig.1 After 48h of culture, the medium(left) showed visible green, indicating that chromoprotein express successfully.]]</th><th></table> | <table><tr><th>[[Image:chromoP.png|thumb|180px|Fig.1 After 48h of culture, the medium(left) showed visible green, indicating that chromoprotein express successfully.]]</th><th></table> | ||
− | + | Growth curve of strains contained this part was detected by spectrophotometer. The values of OD600, OD577 and OD458 were detected and recorded, because OD600 are usually used to show the cell density of <i>E.coli</i> BL21 (DE3), 577 nm is the maximum emission wavelength of gfasPurple chromoprotein and 458 nm is the maximum emission wavelength of amajLime. The detail of the protocol can be viewed in Notebook-Experiment-Growth Curve: | |
<br> | <br> | ||
+ | https://2019.igem.org/Team:XMU-China/Experiments# | ||
+ | <table><tr><th> | ||
+ | [[Image:CPEF.png|thumb|540px|left|Fig.5 The comparison of the value of OD600 between experimental and control groups. PEF is the strain carring <partinfo>BBa_K2922039</partinfo> and <partinfo>BBa_K1033917</partinfo> is the strain carring <partinfo>BBa_K1033917</partinfo> here. +TND/<partinfo>BBa_K1033914</partinfo> is experimental group and TND/<partinfo>BBa_K1033914</partinfo> is control group.]] | ||
+ | </th><th></table> | ||
+ | <br> | ||
For more information, please go to our result: | For more information, please go to our result: | ||
<br> | <br> |
Revision as of 20:40, 21 October 2019
The colicin-E1 operon under pBAD (Arabinose promoter) control with an amajLime chromoprotein reporte
Summary
This is a composite part consisting of an arabinose promoter (BBa_K206000), the CDS of Colicin-E1 (BBa_K2922024), the CDS of Colicin-E1 immunity protein (BBa_K2922025), the CDS of lysis protein (BBa_K2922026) and the amajLime chromoprotein reporter (BBa_K1033914). Each CDS has an RBS (BBa_B0034) behind. pBAD promoter could be induced by arabinose in Escherichia coli BL21 (DE3) strain and then express all proteins mentioned. E coli that can`t express Colicin-E1 immunity protein would be killed by Colicin-E1. Colicin-E1 immunity protein is used to protect itself from attack of extracellular Colicin-E1 and lysis protein helps the release of Colicin-E1. The amajLime chromoprotein reporter is used to present the growth curve specifically of strain which contains this part. This part is constructed in the aim of achieve our "spite" design.
Identification
In order to specifically record the growth curve of strains containing our colicin gene circuit, we combined BBa_K2922037 with amajLime chromoprotein reporter BBa_K1033914 to construct this part. This part was insert into pSB1C3 by standard assembly and transformed into E.coli BL21 (DE3) strain, Colonies were picked and cultured in liquid LB medium.
Growth curve of strains contained this part was detected by spectrophotometer. The values of OD600, OD577 and OD458 were detected and recorded, because OD600 are usually used to show the cell density of E.coli BL21 (DE3), 577 nm is the maximum emission wavelength of gfasPurple chromoprotein and 458 nm is the maximum emission wavelength of amajLime. The detail of the protocol can be viewed in Notebook-Experiment-Growth Curve:
https://2019.igem.org/Team:XMU-China/Experiments#
For more information, please go to our result:
https://2019.igem.org/Team:XMU-China/Results
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 125
Illegal NheI site found at 2360
Illegal NheI site found at 2383 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 65
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 642
Illegal AgeI site found at 2000
Illegal AgeI site found at 2057
Illegal AgeI site found at 2189 - 1000COMPATIBLE WITH RFC[1000]