鸡源沙门氏菌对β-内酰胺类抗生素耐药相关基因的筛选及鉴定

摘要

[目的]从沙门氏菌全基因组范围筛选和鉴定其与β-内酰胺类抗生素耐药性相关的基因.[方法]对临床分离的10株沙门氏菌进行药敏试验,选择耐药性最广泛的菌株作为研究对象,利用mariner转座子对其基因组进行随机突变,获得转座子突变株文库,筛选文库中对β-内酰胺类抗生素敏感的突变体,通过套式PCR、核苷酸测序及序列比对确定突变体中转座子的插入位点及其破坏的基因.[结果]从突变株文库中筛选得到15株分别对青霉素G、氨苄西林和头孢哌酮敏感的突变株,其中3株对三种抗生素都敏感.转座子破坏基因为STM4216 (inner membrane protein)和STM4150 (50S ribosomal protein L1).[结论]筛选出的基因有望成为控制或扭转沙门氏菌耐药性的作用靶点.%[Objective]Genome-wide screening and identification of β-lactam antibiotic resistance genes of Salmonella.[Methods] Drug sensitivity test was performed on clinically separated ten salmonella strains to choose the relatively wide resistance strain as the research object.The mariner transposons was used to construct transposon mutant library of salmonella,and β-1actam antibiotics were used to screen the antibiotics sensitive mutants from the library.Then the transposon insertion sites were determined by nested-PCR,nucleotide sequencing and sequence alignment.[Results] Fifteen strains sensitive topenicillin G,ampicillin,and cefoperazone were selected from the library,and transposoninsertion sites analysis results showed that the damaged genes were STM4216 (inner membrane protein)and STM4150 (50S ribosomal protein L1).[Conclusion] The identified antibiotic resistance genes of salmonella may be supposed to be potential targets for antimicrobial agents.