Difference between revisions of "Part:BBa K2933215"

 
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<partinfo>BBa_K2933215 parameters</partinfo>
 
<partinfo>BBa_K2933215 parameters</partinfo>
 
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==Usage and Biology==
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This composite part is made up with seven basic parts,  the His tag, T7 promoter, RBS ，Linker h ，Linker f，T7 terminatorand and our target protein BcII-194. It encodes a protein which is BcII-194 fused with His tag.Linker h is from the vector pET-28a, which connects the RBS to His tag sequence.Linker f from vector pGEX-6p-1, contains the thrombin restriction site and T7 tag. The fusion protein is about 28.1 kD. In order to gain the highly purified target protein, we add His tag in N-terminal of BcII-194.  It is convenient for us to purify our target protein.<br>
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===Molecular cloning===
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First,we obtained BcII by PCR.<br>
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<p style="text-align: center;">
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[[File:BcII-194 PCR.jpeg|200px|]]<br>
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'''Figure 1.'''    The PCR result of BcII.<br>
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</p>
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==References==
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Sung-Kun Kim, Mara Demuth, Sara R. Schlesinger, Sung Joon Kim, Jonathan Urbanczyk, Robert W. Shaw & Hyunshun Shin (2016) Inhibition of Bacillusanthracis metallo-β- lactamase by compounds with hydroxamic acid functionality, Journal of Enzyme Inhibition and Medicinal Chemistry, 31:sup4, 132-137, DOI: 10.1080/14756366.2016.1222580<br>

Latest revision as of 14:04, 23 September 2019


T7 promoter+RBS b+Linker h+His+Linker f+BCll-194+T7 terminator

This part consists of T7 promoter, RBS and protein coding sequence（His+Linker f+NDM-23），and the biological module can be built into E.coli for protein expression.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 47
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 169
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 47
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 47
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

This composite part is made up with seven basic parts, the His tag, T7 promoter, RBS ，Linker h ，Linker f，T7 terminatorand and our target protein BcII-194. It encodes a protein which is BcII-194 fused with His tag.Linker h is from the vector pET-28a, which connects the RBS to His tag sequence.Linker f from vector pGEX-6p-1, contains the thrombin restriction site and T7 tag. The fusion protein is about 28.1 kD. In order to gain the highly purified target protein, we add His tag in N-terminal of BcII-194. It is convenient for us to purify our target protein.

Molecular cloning

First,we obtained BcII by PCR.

BcII-194 PCR.jpeg
Figure 1. The PCR result of BcII.

References

Sung-Kun Kim, Mara Demuth, Sara R. Schlesinger, Sung Joon Kim, Jonathan Urbanczyk, Robert W. Shaw & Hyunshun Shin (2016) Inhibition of Bacillusanthracis metallo-β- lactamase by compounds with hydroxamic acid functionality, Journal of Enzyme Inhibition and Medicinal Chemistry, 31:sup4, 132-137, DOI: 10.1080/14756366.2016.1222580