[Optimizing Mycobacterium Recombineering System (pJV53) to Promote the Screening of the Mycobacterium Mutants].
| Autor: | Wang C; Laboratory of Infection and Immunity, West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu 610041, China., Ma PJ; Laboratory of Infection and Immunity, West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu 610041, China., Luo T; Laboratory of Infection and Immunity, West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu 610041, China., Bao L; Laboratory of Infection and Immunity, West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu 610041, China. |
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| Jazyk: | čínština |
| Zdroj: | Sichuan da xue xue bao. Yi xue ban = Journal of Sichuan University. Medical science edition [Sichuan Da Xue Xue Bao Yi Xue Ban] 2019 Sep; Vol. 50 (5), pp. 629-634. |
| Abstrakt: | 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 hyg S 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-hyg S 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-hyg S 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. (Copyright© by Editorial Board of Journal of Sichuan University (Medical Science Edition).) |
| Databáze: | MEDLINE |
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