Part:BBa_K2933231
RBS a+Linker g+GST+Linker e+MUS-2
This part consists of RBS a, protein coding sequence(GST+Linker e+MUS-2), the RBS and the protein coding sequence can be connected by linker g. The biological module can be build into E.coli for protein expression. This part can be prefaced with promoters of different strengths and types to regulate expression function.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 113
Usage and Biology
This composite part is made up with five basic parts, the RBSa, the linker g and 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.
References
[1]Al-Bayssari C, Gupta SK, Dabboussi F, Hamze M, Rolain JM. MUS-2, a novel variant of the chromosome-encoded β-lactamase MUS-1, from Myroides odoratimimus. New Microbes New Infect. 2015 Jun 27;7:67-71.
[2] Mammeri H, Bellais S, Nordmann P. Chromosome-encoded beta-lactamases TUS-1 and MUS-1 from Myroides odoratus and Myroides odoratimimus (formerly Flavobacterium odoratum), new members of the lineage of molecular subclass B1 metalloenzymes. Antimicrob Agents Chemother. 2002 Nov;46(11):3561-7.
Molecular cloning
We used the vector pGEX-6p-1 to construct our expression plasmid.
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.
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