ESBLbla SHV-12基因片段的克隆和序列分析  

Cloning and Sequencing of ESBLbla SHV-12 Gene Fragment

作　　者：高虹[1] 马佳毓[1] 张文莉[1] 付英梅[1] 赵月辉[1] 

机构地区：[1]哈尔滨医科大学微生物教研室,哈尔滨黑龙江150086

出　　处：《微生物学杂志》2004年第5期17-19,共3页Journal of Microbiology

基　　金：国家自然科学基金资助项目 ( 3 0 170 816)

摘　　要：分析黑龙江省分离的肺炎克雷伯菌产超广谱 β 内酰胺酶基因片段的序列 ,确定该基因所产ESBLs的亚型。用双纸片法初筛检测产ESBLs菌株 ,聚合酶链反应扩增ESBLs编码基因片段 ,并将其克隆至pUC19载体中 ,双脱氧链终止法测定核苷酸序列以确定亚型。测序结果证实所获得的ESBLs基因片段含10 4 7个核苷酸 ,其基因型为SHV型 ,与SHV 12型ESBLs编码基因相同。上述结果表明 ,黑龙江省内分离的产超广谱 β 内酰胺酶肺炎克雷伯菌含有SHV 12编码基因。The nucleotide sequence of the gene encoding extended spectrum β-lactamases (ESBLs) of Klebsiella pneumoniae isolated from Heilongjiang province was analyzed and the subtype of ESBLs was identified. Double disc test was used to screen the strains of K.pneumoniae producting ESBLs. The genes encoding ESBLs were amplified by polymerase chain reaction (PCR). The target amplification fragment was digested with two restriction endonucleases and then cloned into pUC19 vector. The recombinant vector was sequenced by Sanger's dideoxy chain termination composition method. The sequencing result indicated that the obtained ESBLs gene fragment has 1 047 bp belonging to subtype SHV, which is the same as subtype SHV-12 ESBLs encoding gene. The results of this study revealed the existence of Klebsiella pneumoniae strains containing SHV-12 ESBLs encoding gene.;

关 键 词：肺炎克雷伯菌 超广谱Β-内酰胺酶 聚合酶链反应 克隆 序列分析 

分 类 号：R378.996[医药卫生—病原生物学]