Part:BBa_K2933005
subclass B1 metallo-beta-lactamase MYX-1, codon optimized in E. coli
This part encodes a protein called MYX-1, which is a metallo-beta-lactamase of subclass B1.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 80
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 80
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 80
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 80
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
MYX-1 is a type of subclass B1 metal beta-lactamases, which is found from Myxococcus xanthus. The beta lactamases of subclass B1 family can hydrolyze almost all available beta lactam antibiotics (except aztreonam) clinically, including the broad-spectrum antibiotic carbapenems. Because of the extensive substrate profile of this enzyme, the clinical strains carrying it become a great threat to human life and health.
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 MYX-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.
References
[1]Correction for Goldman et al., Evolution of sensory complexity recorded in a myxobacterial genome. Proc Natl Acad Sci U S A. 2006;103(51):19605. doi:10.1073/pnas.0609567103.
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