Difference between revisions of "Part:BBa K3606813"
 
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<h2>Design:</h2>
 
<h2>Design:</h2>
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Since we encountered difficulties in the expression of McbABCDEFG at the beginning, we decided to check whether the McbA/B/C/D/E/F/G genes can be expressed and translated separately. We successfully cloned McbG into the pGEX plasmid and transcribed it into BL21. After 5 hours of induction of McbG expression with IPTG, the whole protein SDS-PAGE was used to detect whether the McbG product was produced.
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Since we encountered difficulties in the expression of McbABCDEFG at the beginning, we wanted to know whether one promoter can start two consecutive genes. We successfully cloned McbBC into the pGEX plasmid together downstream one promoter and transcribed it into BL21. After 5 hours of induction of McbBC expression with IPTG, the whole protein SDS-PAGE was used to detect whether the McbBC product was produced.  
  
 
<h2>Results:</h2>
 
<h2>Results:</h2>
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https://2020.igem.org/wiki/images/2/22/T--Fudan--img_McbA-E-G-BCD.jpeg
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More information：https://parts.igem.org/Part:BBa_K3606814
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Figure 1. SDS-PAGE result. lane 1: mcbA  lane 2: mcbA+IPTG lane 3: mcbE  lane 4: mcbE+IPTG  lane 5: mcbG  lane 6: mcbG+IPTG  lane 7: mcbBCD  lane 8: mcbBCD+IPTG lane 9: pGEX+ IPTG
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Comparing the 5th and 6th lanes, after IPTG induction, there is a clear band, which is the product of McbG expression.
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 02:41, 5 December 2020


mcbBC

This part is an antibiotic coding gene cluster with proteins for MccB17 maturation.

Design:

Since we encountered difficulties in the expression of McbABCDEFG at the beginning, we wanted to know whether one promoter can start two consecutive genes. We successfully cloned McbBC into the pGEX plasmid together downstream one promoter and transcribed it into BL21. After 5 hours of induction of McbBC expression with IPTG, the whole protein SDS-PAGE was used to detect whether the McbBC product was produced.

Results:

More information：https://parts.igem.org/Part:BBa_K3606814

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]