[Optimizing Mycobacterium Recombineering System (pJV53) to Promote the Screening of the Mycobacterium Mutants].
Sichuan Da Xue Xue Bao Yi Xue Ban
; 50(5): 629-634, 2019 Sep.
Article
in Zh
| MEDLINE
| ID: mdl-31762229
ABSTRACT
OBJECTIVE:
To establish a way for screening Mycobacterium mutants through adding the screening markers into pJV53.METHODS:
The sucrose counter selection gene SacB and mutant hygromycin-resistant gene hygS were inserted into pJV53; The recovery of the hygromycin-resistance indicated the successful homologous recombination in Mycobacterium smegmatis (Ms), which could serve as mutant screening marker; The sucrose counter selection could be used to screen the plasmid-free mutants.RESULTS:
The recombinant plasmid pJV53-SacB-hygS were successfully constructed. The rifampin-resistant rpoB D516Y and rpoB H526Q mutants and MSMEG_4487 G188A mutant were efficiently screened out. All mutants had shed the plasmid successfully.CONCLUSION:
pJV53-SacB-hygS can efficiently contribute to construct and screen the mutants and to get the mutants shedding the plasmid self, which has high value of extensive application; the D516Y and H526Q mutations in gene rpoB of Mycobacterium tuberculosis contribute to its rifampin-resistance.Key words
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Database:
MEDLINE
Main subject:
Mycobacterium smegmatis
/
Drug Resistance, Bacterial
/
Homologous Recombination
/
Mycobacterium tuberculosis
Type of study:
Diagnostic_studies
/
Screening_studies
Language:
Zh
Journal:
Sichuan Da Xue Xue Bao Yi Xue Ban
Year:
2019
Type:
Article
Affiliation country:
China