Part:BBa_K2933190
T7 promoter+RBS b+linker h+His+Linker a+Sumo+Linker b+MUS-2+T7 terminator
The part consists of T7 promoter,RBS and protein coding(His+Linker a+Sumo+Linker b+MUS-2)and the biological module can be built into E.coil for protein expression.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 390
Illegal XbaI site found at 47 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 390
Illegal NheI site found at 167
Illegal NheI site found at 1238 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 390
Illegal BglII site found at 279
Illegal BamHI site found at 478 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 390
Illegal XbaI site found at 47 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 390
Illegal XbaI site found at 47 - 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
This composite part is made up with nine basic parts, T7 promoter, the RBS b， the linker h， His tag，the linker a, Sumo tag, linker b, the gene of MUS-2 and T7 terminator.It encodes a protein which is MUS-2 fused with His and Sumo tag. The fusion protein is about 40.3 kD. In order to gain the highly purified target protein, we add GST tag in N-terminal of MUS-2 and combine Sumo tag to increased protein solubility. 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 pET28B-Sumo to construct our expression plasmid.
Figure 1. The PCR result of MUS-2.
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.| None |