肝螺杆菌鞭毛蛋白B基因的克隆和序列测定  

Cloning and Sequence Analysis of the Flagellin B Gene of Helicobacter hepaticus

作　　者：高正琴[1] 张强[2] 贺争鸣[1] 邢进[1] 岳秉飞[1] 冯育芳[1] 

机构地区：[1]中国药品生物制品检定所,北京100050 [2]首都医科大学附属北京佑安医院,北京100069

出　　处：《实验动物与比较医学》2008年第4期220-224,229,共6页Laboratory Animal and Comparative Medicine

摘　　要：目的对肝螺杆菌BJ 0801鞭毛蛋白B(flagellin B,fla B)基因进行扩增、克隆和序列测定,以探讨其功能。方法根据已公布的肝螺杆菌ATCC 51449序列,设计合成一对引物,以肝螺杆菌BJ 0801株DNA为模板,扩增fla B基因片段,与pGEM-T载体连接并转化感受态E.coliDH5α,提取的重组质粒经双酶切、PCR、电泳鉴定并进行序列测定。结果目的基因片段长度为1 545 bp,与已公布的肝螺杆菌ATCC 51449核苷酸同源性达99%。结论成功克隆出肝螺杆菌BJ 0801鞭毛蛋白B基因序列,为进一步研究该基因的功能奠定了基础。Objective The flagellin B （fla B） gene ofHelicobacter hepaticus （H. hepaticus） was amplified, cloned and sequenced in order to investigate its function. Method A pair of primer was used for amplifying H. hepaticus fla B gene of BJ 0801 strain with PCR. The amplified fragment was cloned into the pGEM-T vector and transformed into competence E. coli DH5α. The recombinant plasmid was identified by PCR, endonuclease analysis and electrophoresis, and confirmed by sequencing. The nucleotide sequence was compared with thefla B gene of H, hepaticus ATCC 51449. Results The results demonstrated that nucleotide homology of H, hepaticus BJ 0801 fla B gene with H. hepaticus ATCC 51449 fla B gene was 99%. Conclusion Characterization of the H. hepaticus BJ 0801 strain at the genetic identification would be an important step toward investigating H. hepaticusfla B gene function.

关 键 词：肝螺杆菌 鞭毛蛋白 克隆 测序 

分 类 号：Q93[生物学—微生物学] Q78