Difference between revisions of "Part:BBa K2933174"

 
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<partinfo>BBa_K2933174 parameters</partinfo>
 
<partinfo>BBa_K2933174 parameters</partinfo>
 
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===Usage and Biology===
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<br>
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===Molecular cloning===
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First, we used the vector pGEX-6p-1 and pET-28b_SUMO to construct our expression plasmid. And then we converted the plasmid constructed to ''E. coli'' DH5α to expand the plasmid largely.<br>
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<p style="text-align: center;">
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  [[File:TMB-2-PCR.png|400px]]<br>
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'''Figure 1.'''  Left: The PCR result of TMB-2. Right: The verification results by enzyme digestion.<br>
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</p>
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After verification, it was determined that the construction is successful. We converted the plasmid to ''E. coli'' BL21(DE3) for expression and purification.<br>

Revision as of 12:44, 21 September 2019


Tac promoter+RBS a+Linker g+GST+Linker e+TMB-2

This part consists of Tac promoter,RBS and protein coding sequence (GST+Linker e+TMB-2),and the biological module can be built into E.coli for protein expression.


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 181


Usage and Biology


Molecular cloning

First, we used the vector pGEX-6p-1 and pET-28b_SUMO to construct our expression plasmid. And then we converted the plasmid constructed to E. coli DH5α to expand the plasmid largely.

TMB-2-PCR.png
Figure 1. Left: The PCR result of TMB-2. Right: The verification results by enzyme digestion.

After verification, it was determined that the construction is successful. We converted the plasmid to E. coli BL21(DE3) for expression and purification.