Difference between revisions of "Part:BBa K2922039"
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===Summary=== | ===Summary=== | ||
| − | This is a composite part consisting of an arabinose promoter (<partinfo>BBa_K206000</partinfo>), the CDS of Colicin-E1 (<partinfo>BBa_K2922024</partinfo>), the CDS of Colicin-E1 immunity protein (<partinfo>BBa_K2922025</partinfo>), the CDS of lysis protein (<partinfo>BBa_K2922026</partinfo>) and the amajLime chromoprotein reporter (<partinfo>BBa_K1033914</partinfo>). Each CDS has an RBS (<partinfo>BBa_B0034</partinfo>) behind. pBAD promoter could be induced by arabinose in <i>Escherichia coli</i> BL21 (DE3) strain and then express all proteins mentioned. <i>E coli</i> that can`t express Colicin-E1 immunity protein would be killed by Colicin-E1 | + | This is a composite part consisting of an arabinose promoter (<partinfo>BBa_K206000</partinfo>), the CDS of Colicin-E1 (<partinfo>BBa_K2922024</partinfo>), the CDS of Colicin-E1 immunity protein (<partinfo>BBa_K2922025</partinfo>), the CDS of lysis protein (<partinfo>BBa_K2922026</partinfo>) and the amajLime chromoprotein reporter (<partinfo>BBa_K1033914</partinfo>). Each CDS has an RBS (<partinfo>BBa_B0034</partinfo>) behind. pBAD promoter could be induced by arabinose in <i>Escherichia coli</i> BL21 (DE3) strain and then express all proteins mentioned. <i>E coli</i> 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. |
<table><tr><th>[[Image:E1design.png|thumb|800px|The diagram of colicin-E1 kit design.]]</th><th></table> | <table><tr><th>[[Image:E1design.png|thumb|800px|The diagram of colicin-E1 kit design.]]</th><th></table> | ||
Revision as of 14:20, 19 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, Clonies were picked and cultured in liquid LB medium.
Fig.1 After 48h of culture, the medium showed visible green, indicating that chromoprotein express successfully.
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]