| Autor: |
DI Y; State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China., Bai J; State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China., Chi M; State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China., Fan W; Laboratory of Zoonosis, China Animal Health and Epidemiology Center, Qingdao 266032, Shangdong, China., Zhang X; State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China. |
| Abstrakt: |
As the only translational factor that plays a critical role in two translational processes (elongation and ribosome regeneration), GTPase elongation factor G (EF-G) is a potential target for antimicrobial agents. Both Mycobacterium smegmatis and Mycobacterium tuberculosis have two EF-G homologous coding genes, M sm EFG1 ( MSMEG_1400 ) and Msm EFG2 ( MSMEG_6535 ), fusA1 ( Rv0684 ) and fusA2 ( Rv0120c ), respectively. Msm EFG1 ( MSMEG_1400 ) and fusA1 ( Rv0684 ) were identified as essential genes for bacterial growth by gene mutation library and bioinformatic analysis. To investigate the biological function and characteristics of EF-G in mycobacterium, two induced EF-G knockdown strains (Msm-ΔEFG1(KD) and Msm-ΔEFG2(KD)) from Mycobacterium smegmatis were constructed by clustered regularly interspaced short palindromic repeats interference (CRISPRi) technique. EF-G2 knockdown had no effect on bacterial growth, while EF-G1 knockdown significantly retarded the growth of mycobacterium, weakened the film-forming ability, changed the colony morphology, and increased the length of mycobacterium. It was speculated that EF-G might be involved in the division of bacteria. Minimal inhibitory concentration assay showed that inhibition of EF-G1 expression enhanced the sensitivity of mycobacterium to rifampicin, isoniazid, erythromycin, fucidic acid, capreomycin and other antibacterial agents, suggesting that EF-G1 might be a potential target for screening anti-tuberculosis drugs in the future. |
