Difference between revisions of "Part:BBa K3606812"
 
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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.
+
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, yet the result of colony PCR turns out as false positive.
  
 
<h2>Results:</h2>
 
<h2>Results:</h2>
  
−
We've constructed the plasmid for mcbB and mcbD expression, yet the results of colony PCR is false positive thus further experiment for the fundamental validation of the antimicrobial peptide(mccb17) expression is needed.
+
We've constructed the plasmid for mcbG expression, yet the results of colony PCR is false positive thus further experiment is needed for the fundamental validation of the antimicrobial peptide(mccb17) expression.
 
https://2020.igem.org/wiki/images/f/f3/T--Fudan--Poster_Mcb-gels.jpg
 
https://2020.igem.org/wiki/images/f/f3/T--Fudan--Poster_Mcb-gels.jpg
−
Figure1. gel map for mcbA, mcbC, mcbE and mcbF
+
Figure2. gel map for mcbA, mcbC, mcbE and mcbF
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 02:40, 5 December 2020


mcbG

Usage and Biology:

McbG is responsible for immunity against MccB17.

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 McbG expression with IPTG, the whole protein SDS-PAGE was used to detect whether the McbG product was produced, yet the result of colony PCR turns out as false positive.

Results:

We've constructed the plasmid for mcbG expression, yet the results of colony PCR is false positive thus further experiment is needed for the fundamental validation of the antimicrobial peptide(mccb17) expression. T--Fudan--Poster_Mcb-gels.jpg Figure2. gel map for mcbA, mcbC, mcbE and mcbF

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]