Difference between revisions of "Part:BBa K2933017"

 
Line 17: Line 17:
 
<partinfo>BBa_K2933017 parameters</partinfo>
 
<partinfo>BBa_K2933017 parameters</partinfo>
 
<!-- -->
 
<!-- -->
 +
===Usage and Biology===
 +
RST-1 is a type of subclass B1 metal beta-lactamases. 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.<br>
 +
==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.<br>
 +
<p style="text-align: center;">
 +
  [[File:T--TJUSLS China--PST-1 PCRmeiqie.png|300px]]  [[File:T--TJUSLS China--PST-1 meiqie.png|300px]]<br>
 +
'''Figure 1.'''  Left: The PCR result of PST-1. Right: The verification results by enzyme digestion.<br>
 +
</p>
 +
After verification, it was determined that the construction is successful. We converted the plasmid to ''E. coli'' BL21(DE3) for expression and purification.<br>

Revision as of 08:03, 15 September 2019


subclass B1 metallo-beta-lactamase PST-1, codon optimized in E. coli

This part encodes a protein called PST-1, which is a metallo-beta-lactamase of subclass B1.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

RST-1 is a type of subclass B1 metal beta-lactamases. 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.

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.

300px 300px
Figure 1. Left: The PCR result of PST-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.