Part:BBa_K2933019

subclass B1 metallo-beta-lactamase BlaB-14, codon optimized in E. coli

This part encodes a protein called BlaB-14, which is a metallo-beta-lactamase of subclass B1.

BlaB is a chromosome-encoded MBL produced by Elizabethkingia meningoseptica. It has a broad substrate profile and is produced constitutively. C. meningosepticum is an ubiquitous Gramnegative rod bacterium of clinical relevance because it can cause neonatal meningitis, adult septicemia, and nosocomial infections and is resistant to most β-lactams, including carbapenems. This pathogen's multidrug-resistant phenotype and attendant lack of adequate molecular mechanistic data limit the current approaches for its effective management in hospitals and public health settings

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 561
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Usage and Biology

BlaB-14 is a type of subclass B metal beta-lactamases. The beta lactamases of the blaB 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.

References

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. (a) The PCR result of BlaB-14. (b) The verification results by enzyme digestion.