Coding

Part:BBa_K2933143

Designed by: Ruihan Dong   Group: iGEM19_TJUSLS_China   (2019-09-14)


His+Linker f+beta-lactamase II [Erythrobacter litoralis HTCC2594]

This part encodes the fusion protein of His tag and beta-lactamase II to promote the expression and purification of beta-lactamase II [Erythrobacter litoralis HTCC2594].

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 51
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 570
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

This composite part is made up with three basic parts, the HiS tag, the thrombin restriction site and our target protein ElblaII. It encodes a protein which is ElblaII fused with His tag. The fusion protein is about 27.5 kD. It is convenient for us to purify our target protein.

Molecular cloning

First, we used the vector pET28a to construct our expression plasmid. And then we converted the plasmid constructed to E. coli DH5α to expand the plasmid largely.

TJUSLS China--Elbla2-1-PCR.png
Figure 1. The PCR result of ElblaII.

References

[1] Girlich D, Poirel L, Nordmann P, Diversity of naturally occurring Ambler class B metallo-β-lactamases in Erythrobacter spp. The Journal of Antimicrobial Chemotherapy [31 Jul 2012, 67(11):2661-2664]

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