Difference between revisions of "Part:BBa K2933107"
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<partinfo>BBa_K2933107 short</partinfo> | <partinfo>BBa_K2933107 short</partinfo> | ||
| − | This part encodes the fusion protein of GST tag and | + | This part encodes the fusion protein of GST tag and MYX-1 to promote the expression and purification of target protein(MYX-1). |
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===Usage and Biology=== | ===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 | + | This composite part is made up with three basic parts, the GST tag, the cutting site of Prescission Protease and our target protein MYX-1. It encodes a protein which is MYX-1 fused with GST tag. The fusion protein is about 53.9 kD. In order to gain the highly purified target protein, we add GST tag in N-terminal of MYX-1 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> |
===Molecular cloning=== | ===Molecular cloning=== | ||
First, we used the vector pGEX-6p-1 to construct our expression plasmid. And then we converted the plasmid constructed to ''E. coli'' DH5α to expand the plasmid largely.<br> | First, we used the vector pGEX-6p-1 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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After verification, it was determined that the construction is successful. We converted the plasmid to ''E. coli'' BL21(DE3) for expression and purification.<br> | After verification, it was determined that the construction is successful. We converted the plasmid to ''E. coli'' BL21(DE3) for expression and purification.<br> | ||
| + | ===References=== | ||
| + | [1]Correction for Goldman et al., Evolution of sensory complexity recorded in a myxobacterial genome. Proc Natl Acad Sci U S A. 2006;103(51):19605. doi:10.1073/pnas.0609567103.<br> | ||
Latest revision as of 14:54, 23 September 2019
GST+Linker+MYX-1
This part encodes the fusion protein of GST tag and MYX-1 to promote the expression and purification of target protein(MYX-1).
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 778
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 778
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 778
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 778
- 1000INCOMPATIBLE 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 MYX-1. It encodes a protein which is MYX-1 fused with GST tag. The fusion protein is about 53.9 kD. In order to gain the highly purified target protein, we add GST tag in N-terminal of MYX-1 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
First, we used the vector pGEX-6p-1 to construct our expression plasmid. And then we converted the plasmid constructed to E. coli DH5α to expand the plasmid largely.
Figure 1. Left: The PCR result of MYX-1. 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.
References
[1]Correction for Goldman et al., Evolution of sensory complexity recorded in a myxobacterial genome. Proc Natl Acad Sci U S A. 2006;103(51):19605. doi:10.1073/pnas.0609567103.