利用反义技术抑制大肠杆菌β-内酰胺酶基因表达的实验研究  

Down-regulation of β-lactamase TEM-1 Gene expression by antisense RNA expressing plasmid in E.coli

作　　者：杨爱琼[1] 谢勇恩[2] 

机构地区：[1]川北医学院附属医院药剂科,四川南充637000 [2]川北医学院病理生理学教研室,四川南充637007

出　　处：《公共卫生与临床医学》2009年第4期252-254,共3页Public health and dinical medicine

基　　金：四川省教育厅高校重点实验室专项经费资助项目（No：2006ZD021）

摘　　要：目的探讨利用反义表达质粒抑制细菌耐药基因表达的可行性。方法根据GenBank中β-内酰胺酶TEM-1基因序列设计一对寡核苷酸5J物，通过PCR扩增获取TEM-1基因，将其反向克隆入质粒pBK—CMV的多克隆位点构建反义表达质粒，将此反义表达质粒导入携带TEM-1型β-内酰胺酶的大肠杆菌，Western—blotting检测TEM-1表达情况。结果成功构建了TEM—1基因的反义表达质粒，将其导入携带TEM-1型β-内酰胺酶的大肠杆菌后，Western—blotting检测发现TEM-1表达显著下调。结论利用反义表达质粒可抑制大肠杆菌β-内酰胺酶TEM-1基因表达。Objective To determine the feasibility of introducing antisense RNA expressing plasmid to suppress the expression of the drug-resistant gene in E. coll. Methods TEM-1 gene fragments were generated by polymerase chain reaction （PCR）. A recombinant plasmid was constructed by inserted the TEM- 1 gene into the multiple cloning site of pBK-CMV vector in the antisense direction. The recombinant plasmid was transformed into E.coli carrying TEM-1 gene. TEM-1 expression was determined by Western-blotting. Results A recombinant plasmid expressing antisense TEM-1 RNA was successfully constructed. TEM-1 gene expression was inhibited by the antisense RNA expressing plasmid in E.coli. Conclusions Antisense technique has potential as novel strategy for blocking the expression of drug-resistant gene in E.coli.

关 键 词：大肠杆菌 TEM-1基因 反义表达质粒 

分 类 号：R378.21[医药卫生—病原生物学]