Part:BBa_K2933273

RBS+Linker h+His+Linker f+MUS-2+T7 terminator

This part consists of RBS, Linker h, protein coding sequence(His+Linker f+MUS-2) and T7 terminator,and 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

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 93
  Illegal NheI site found at 877
  
• 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 six basic parts(RBS , Linker h, His, Linker f , MUS-2 and T7 terminator ). It encodes a protein which is MUS-2 fused with His tag. The fusion protein is about 28.3 kD. It is convenient for us to purify our target protein.

Molecular cloning

We used the vector pET28a 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.