Difference between revisions of "Part:BBa K2933008"
 
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===Usage and Biology===
 
===Usage and Biology===
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To be uploaded
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blaPEDO-1, a new carbapenem-hydrolyzing beta-lactamases, this enzyme has not yet emerged in clinical settings but constitute potential carbap- enem resistance determinants in pathogenic bacterial species, as demonstrated by their ability to confer resistance to ampi- cillin and various cephalosporins, as well as reduced suscepti- bility to carbapenems, once expressed in E. coli.
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===References===
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[1]Dereje Dadi Gudeta, Valeria Bortolaia,The Soil Microbiota Harbors a Diversity of Carbapenem-Hydrolyzing β-Lactamases of Potential Clinical Relevance[J]，Antimicrobial Agents and chemotherapy,January 2016<br>
 
===Molecular cloning===
 
===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>
 
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;">
 
<p style="text-align: center;">
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   [[File:PEDO-1-PCR.png|500px]]<br>
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   [[File:PEDO-1-PCR.png|400px]][[File:PEDO-1-enzyme digestion.png|400px]]<br>
 
'''Figure 1.'''  Left: The PCR result of PEDO-1. Right: The verification results by enzyme digestion.<br>
 
'''Figure 1.'''  Left: The PCR result of PEDO-1. Right: The verification results by enzyme digestion.<br>
 
</p>
 
</p>

Latest revision as of 12:07, 23 September 2019


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

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


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 537
    Illegal PstI site found at 823
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 537
    Illegal PstI site found at 823
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 537
    Illegal PstI site found at 823
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 537
    Illegal PstI site found at 823
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

blaPEDO-1, a new carbapenem-hydrolyzing beta-lactamases, this enzyme has not yet emerged in clinical settings but constitute potential carbap- enem resistance determinants in pathogenic bacterial species, as demonstrated by their ability to confer resistance to ampi- cillin and various cephalosporins, as well as reduced suscepti- bility to carbapenems, once expressed in E. coli.

References

[1]Dereje Dadi Gudeta, Valeria Bortolaia,The Soil Microbiota Harbors a Diversity of Carbapenem-Hydrolyzing β-Lactamases of Potential Clinical Relevance[J]，Antimicrobial Agents and chemotherapy,January 2016

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.

PEDO-1-PCR.pngPEDO-1-enzyme digestion.png
Figure 1. Left: The PCR result of PEDO-1. Right: The verification results by enzyme digestion.