Difference between revisions of "Part:BBa K2933142"

 
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<partinfo>BBa_K2933142 parameters</partinfo>
 
<partinfo>BBa_K2933142 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 SPG-1. It encodes a protein which is SPG-1 fused with His tag. The fusion protein is about 30.2 kD. It is convenient for us to purify our target protein.
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===Molecular cloning===
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First, we used the vector pET28a 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:SPG-PCR.png]]<br>
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'''Figure 1.'''  a: The PCR result of SPG. b: 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 04:09, 22 September 2019


His+Linker f+SPG-1

This part encodes the fusion protein of His tag and SPG-1 to promote the expression and purification of target protein(SPG-1).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 51
  • 21
    COMPATIBLE WITH RFC[21]
  • 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 SPG-1. It encodes a protein which is SPG-1 fused with His tag. The fusion protein is about 30.2 kD. It is convenient for us to purify our target protein.

Molecular cloning

First, we used the vector pET28a to construct our expression plasmid. And then we converted the plasmid constructed to E. coli DH5α to expand the plasmid largely.

SPG-PCR.png
Figure 1. a: The PCR result of SPG. b: 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.