title: [Establishment and drug susceptibility test of isoniazid resistant Mycobacterium smegmatis].
abstract: With the emergence of drug resistant tuberculosis, it is very urgent to find novel anti-tuberculosis drugs, especially novel anti-drug-resistant tuberculosis drugs. Because of the slow growth and the need to work in a biosafty environment of Mycobacterium tuberculosis, the development of evaluation of drug effect is severely impeded. In order to solve these issues, non-pathogenic fast-growing Mycobacterium smegmatis is introduced as test organism. The inhA is one of a target of isoniazid (INH) overexpression or mutation of this gene in Mycobacterium tuberculosis conferring resistant to INH. A recombinant plasmid bearing inhA was constructed and electroporated into Mycobacterium smegmatis, using shuttle expression vector pMV261. Transformants were induced to express a protein of inhA, identified by SDS-PAGE. Results show that Mycobacterium smegmatis containing inhA plasmids exhibited 100-fold or greater increased resistance to INH, but it conferred no increased resistance to others first-line anti-tuberculosis drugs. Resazurin microtiter assay plate testing of Mycobacterium smegmatis susceptibility to drugs is a rapid, simple, and inexpensive method and could decrease color background of drugs by detecting fluorescence. It will be benefit for high-throughout screening of drugs of anti-isoniazid-resistant Mycobacteria.
authors.affiliation: Laboratory of Immunobiology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing 100050, China.
authors.forename: Ping-ping
authors.initials: PP
authors.lastname: Jia
authors.forename: Li-li
authors.initials: LL
authors.lastname: Zhao
authors.forename: Xiao-yu
authors.initials: XY
authors.lastname: Li
authors.forename: Quan
authors.initials: Q
authors.lastname: Zhang
authors.forename: Zhen-long
authors.initials: ZL
authors.lastname: Liu
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.forename: Li-yan
authors.initials: LY
authors.lastname: Yu
authors.forename: Li-xun
authors.initials: LX
authors.lastname: Zhao
authors.forename: Shan
authors.initials: S
authors.lastname: Cen
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000904:Antibiotics, Antitubercular / Q000494:pharmacology; D000995:Antitubercular Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D024881:Drug Resistance, Bacterial; D018274:Electroporation; D004977:Ethambutol / Q000494:pharmacology; D007538:Isoniazid / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D020102:Mycobacterium smegmatis / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D012293:Rifampin / Q000494:pharmacology; D013307:Streptomycin / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D000904:Antibiotics, Antitubercular; D000995:Antitubercular Agents; D001426:Bacterial Proteins; D004977:Ethambutol; D010088:Oxidoreductases; C085257:InhA protein, Mycobacterium; D007538:Isoniazid; D012293:Rifampin; D013307:Streptomycin