Difference between revisions of "Part:BBa K2933273"
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===Usage and Biology===
 
===Usage and Biology===
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This composite part is made up with four basic parts(Tac promoter，RBS a , Linker g , GST+Linker e+MUS-2). 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.
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This composite part is made up with four basic parts(RBS，Linker h , T7 terminator and His+Linker f+MUS-2). 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===
 
===Molecular cloning===

Revision as of 07:36, 24 September 2019


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

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

MUS PCR.png
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.