Difference between revisions of "Part:BBa K3606806"
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<partinfo>BBa_K3606806 short</partinfo>
 
<partinfo>BBa_K3606806 short</partinfo>
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<h2>Usage and Biology:</h2>
 
McbA is the structural gene of MccB17, which is produced by the gene cluster of McbABCDEFG.  
 
McbA is the structural gene of MccB17, which is produced by the gene cluster of McbABCDEFG.  
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<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 McbA expression with IPTG, the whole protein SDS-PAGE was used to detect whether the McbA product was produced.
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<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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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 1th and 2th lanes, the difference is small. This may be due to the low expression or low molecular weight of McbA products.
  
 
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Revision as of 01:40, 28 October 2020


mcbA

Usage and Biology:

McbA is the structural gene of MccB17, which is produced by the gene cluster of McbABCDEFG.

Design:

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 McbA expression with IPTG, the whole protein SDS-PAGE was used to detect whether the McbA product was produced.

Results:

T--Fudan--img_McbA-E-G-BCD.jpeg

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

Comparing the 1th and 2th lanes, the difference is small. This may be due to the low expression or low molecular weight of McbA products.

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]