Difference between revisions of "Part:BBa K2933155"

 
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<partinfo>BBa_K2933155 parameters</partinfo>
 
<partinfo>BBa_K2933155 parameters</partinfo>
 
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===Usage and Biology===
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This composite part is made up with three basic parts, the HiS tag, the thrombin restriction site and our target protein VIM-66. It encodes a protein which is VIM-66 fused with His tag. The fusion protein is about 28.3 kD. It is convenient for us to purify our target protein.
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===Molecular cloning===
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We insert VIM-66 gene into the standard vector then transfer it into E.coli.
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  [[File:VIM-66-PCR.jpeg|600px|center|]] 
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<p style="text-align: center;">
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'''Figure 1.''' Left: The result of PCR, Right:The result of double enzyme digestion verification.LaneM,Marker, Lane1, the plasmid with VIM-66, Lane2, after double enzyme verification

Revision as of 07:56, 22 September 2019


His+Linker f+VIM-66

This part encodes the fusion protein of His tag and VIM-66 to promote the expression and purification of target protein(VIM-66).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 51
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 824
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

This composite part is made up with three basic parts, the HiS tag, the thrombin restriction site and our target protein VIM-66. It encodes a protein which is VIM-66 fused with His tag. The fusion protein is about 28.3 kD. It is convenient for us to purify our target protein.

Molecular cloning

We insert VIM-66 gene into the standard vector then transfer it into E.coli.

VIM-66-PCR.jpeg

Figure 1. Left: The result of PCR, Right:The result of double enzyme digestion verification.LaneM,Marker, Lane1, the plasmid with VIM-66, Lane2, after double enzyme verification