Difference between revisions of "Part:BBa K2933111"

 
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<partinfo>BBa_K2933111 parameters</partinfo>
 
<partinfo>BBa_K2933111 parameters</partinfo>
 
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===Usage and Biology===
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This composite part is made up with three basic parts, the GST tag, the cutting site of Prescission Protease and our target protein MUS-2. It encodes a protein which is MUS-2 fused with GST tag. The fusion protein is about 54.3 kD. In order to gain the highly purified target protein, we add GST tag in N-terminal of MUS-2 and combine the two parts with the cutting site of Prescission Protease. The fusion protein can be cut off at the cutting site by Prescission Protease. It is convenient for us to purify our target protein.<br>
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===Molecular cloning===
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We used the vector pGEX-6p-1 to construct our expression plasmid.
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<p style="text-align: center;">
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  [[File:TJUSLS China--MUS-2-PCR.png|500px]]<br>
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'''Figure 1.'''  Left: The PCR result of MUS-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.

Revision as of 15:53, 14 September 2019


GST+Linker+MUS-2

This part encodes the fusion protein of GST tag and MUS-2 to promote the expression and purification of target protein(MUS-2).

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 85


Usage and Biology

This composite part is made up with three basic parts, the GST tag, the cutting site of Prescission Protease and our target protein MUS-2. It encodes a protein which is MUS-2 fused with GST tag. The fusion protein is about 54.3 kD. In order to gain the highly purified target protein, we add GST tag in N-terminal of MUS-2 and combine the two parts with the cutting site of Prescission Protease. The fusion protein can be cut off at the cutting site by Prescission Protease. It is convenient for us to purify our target protein.

Molecular cloning

We used the vector pGEX-6p-1 to construct our expression plasmid.

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

After verification, it was determined that the construction is successful.