Difference between revisions of "Part:BBa K3034000"

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===References===
 
===References===
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[1]Chávez-Jacobo, Víctor M., et al. "CrpP is a novel ciprofloxacin-modifying enzyme encoded by the Pseudomonas aeruginosa pUM505 plasmid." Antimicrobial agents and chemotherapy 62.6 (2018): e02629-17.
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[1]Chávez-Jacobo, Víctor M., et al. ''CrpP is a novel ciprofloxacin-modifying enzyme encoded by the Pseudomonas aeruginosa pUM505 plasmid.'' Antimicrobial agents and chemotherapy 62.6 (2018): e02629-17.
  
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[2]Foucout, Léna?g, et al. "Synthesis, radiosynthesis and biological evaluation of 1, 4-dihydroquinoline derivatives as new carriers for specific brain delivery." Organic & biomolecular chemistry 7.18 (2009): 3666-3673.
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[2]Foucout, et al. ''Synthesis, radiosynthesis and biological evaluation of 1, 4-dihydroquinoline derivatives as new carriers for specific brain delivery.'' Organic & biomolecular chemistry 7.18 (2009): 3666-3673.
 
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K3034000 parameters</partinfo>
 
<partinfo>BBa_K3034000 parameters</partinfo>
 
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Revision as of 15:26, 11 October 2019

CrpP：Ciprofloxacin-Modifying Enzyme

CrpP is a novel ciprofloxacin-modifying enzyme which can phosphorylate ciprofloxacin(CIP). Then the phosphorylated CIP goes through multiple steps of degradation spontaneously and produces 1,4-dihydroquinoline finally[1]. 1,4-dihydroquinoline is used as a kind of new carriers for specific brain delivery [2], so we can infer that it's nontoxic. This part is responsible for degradation of ciprofloxacin in our project.

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]

Characterization

Molecular weight

This gene codes for a protein of 65 amino acids with a molecular mass about 15 kDa.

Expression of CrpP-Histag

Expression of CrpP-Histag is unsatisfying in E.coli DH5α, so we clone pelB-5D, CrpP and 6×Histag CDS into an expression vector, then the vector is transformed into E.coli BL21(DE3). We use IPTG to induce its overexpression and SDS-PAGE is used to detect target protein's expression.

Fig. 1SDS-PAGE of CrpP-Histag. Lane 1: Protein ladder; Lane 2、3、5 and 6: experimental group(0.5mM IPTG is added); Lane 3、4、7 and 8: control group(without 0.5mM IPTG)

Enzyme activity of CrpP

References

[1]Chávez-Jacobo, Víctor M., et al. CrpP is a novel ciprofloxacin-modifying enzyme encoded by the Pseudomonas aeruginosa pUM505 plasmid. Antimicrobial agents and chemotherapy 62.6 (2018): e02629-17.

[2]Foucout, et al. Synthesis, radiosynthesis and biological evaluation of 1, 4-dihydroquinoline derivatives as new carriers for specific brain delivery. Organic & biomolecular chemistry 7.18 (2009): 3666-3673.