Huang Li, Yuan Hui, Liu Ma-Feng, Zhao Xin-Xin, Wang Ming-Shu, Jia Ren-Yong, Chen Shun, Sun Kun-Feng, Yang Qiao, Wu Ying, Chen Xiao-Yue, Cheng An-Chun, Zhu De-Kang
Research Center of Avian Diseases, College of Veterinary Medicine of Sichuan Agricultural UniversityChengdu, China; Key Laboratory of Animal Disease and Human Health of Sichuan ProvinceChengdu, China.
Key Laboratory of Animal Disease and Human Health of Sichuan ProvinceChengdu, China; Institute of Preventive Veterinary Medicine, Sichuan Agricultural UniversityChengdu, China.
Front Microbiol. 2017 Mar 1;8:297. doi: 10.3389/fmicb.2017.00297. eCollection 2017.
causes serositis and septicaemia in domestic ducks, geese, and turkeys. Traditionally, the antibiotics were used to treat this disease. Currently, our understanding of susceptibility to chloramphenicol and the underlying resistance mechanism is limited. In this study, the gene was identified in 69/192 (36%) isolated from different regions in China, including CH-2 that has been sequenced in previous study. Sequence analysis suggested that there are two copies of gene in this strain. Only both two copies of the mutant strain showed a significant decrease in resistance to chloramphenicol, exhibiting 4 μg/ml in the minimum inhibitory concentration for this antibiotic, but not for the single gene deletion strains. Functional analysis of the gene via expression in BL21 (DE3) cells and site-directed mutagenesis indicated that His79 is the main catalytic residue of CAT in . These results suggested that chloramphenicol resistance of CH-2 is mediated by the genes. Finally, homology analysis of types A and B CATs indicate that comprises type B3 CATs.
在家鸭、鹅和火鸡中引起浆膜炎和败血症。传统上,抗生素被用于治疗这种疾病。目前,我们对氯霉素敏感性及其潜在耐药机制的了解有限。在本研究中,在中国不同地区分离的192株菌株中有69株(36%)鉴定出该基因,其中包括先前研究中已测序的CH-2。序列分析表明,该菌株中有两个该基因的拷贝。只有两个拷贝的该基因突变菌株对氯霉素的耐药性显著降低,该抗生素的最低抑菌浓度为4μg/ml,而单基因缺失菌株则没有。通过在BL21(DE3)细胞中表达和定点诱变对该基因进行功能分析表明,His79是该基因中氯霉素乙酰转移酶(CAT)的主要催化残基。这些结果表明,CH-2对氯霉素的耐药性是由该基因介导的。最后,对A类和B类CATs的同源性分析表明,该基因属于B3类CATs。