Part:BBa_K2933012
subclass B1 metallo-beta-lactamase JOHN-1, codon optimized in E. coli
This part encodes a protein called JOHN-1, which is a metallo-beta-lactamase of subclass B1.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 76
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 76
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 76
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 76
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 600
Usage and Biology
Flavobacterium johnsoniae (formerly Cytophaga johnsonae) is an environmental bacterium that can cause skin lesions in fish and is a plant pathogen.
Origin(organism)
Klebsiella pneumoniae.
References
[1]Naas Thierry,Bellais Samuel,Nordmann Patrice. Molecular and biochemical characterization of a carbapenem-hydrolysing beta-lactamase from Flavobacterium johnsoniae.[J]. Journal of Antimicrobial Chemotherapy,2003,51(2).
Molecular cloning
First, we used the vector pGEX-6p-1 to construct our expression plasmid. And then we converted the plasmid constructed to E. coli DH5α to expand the plasmid largely.
Figure 1. Left: The PCR result of JOHN-1. Right: The verification results by enzyme digestion.
After verification, it was determined that the construction is successful. We converted the plasmid to E. coli BL21(DE3) for expression and purification.
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