编码日本血吸虫大陆株磷酸丙糖异构酶(TPI)抗原基因的克隆与测序  

CLONING AND SEQUENCING OF THE GENE ENCODING TPI ANTIGEN FROM CHINESE STRAIN OF SCHISTOSOMA JAPONICUM

作　　者：缪应新[1] 刘述先[2] 

机构地区：[1]华东医院上海市老年医学研究所,上海200040 [2]中国预防医学科学院寄生虫病研究所,上海200025

出　　处：《中国寄生虫学与寄生虫病杂志》1997年第6期349-352,共4页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：世界卫生组织热带病研究与培训专门规划司的资助

摘　　要：目的 :获得编码日本血吸虫大陆株磷酸丙糖异构酶 ( Sjc TPI)的 c DNA基因片段 ,对其克隆与测序。方法 :以日本血吸虫大陆株成虫总 RNA为模板 ,经过逆转录 -聚合酶链 (式 )反应( RT- PCR)获得编码 Sjc TPI抗原的 c DNA片段 ,并克隆于载体 M13mp18、M13mp19,双脱氧链末端终止法测 DNA序列。结果 :体外 PCR扩增获得了编码 Sjc TPI的 0 .75kb c DNA片段 ,6个重组克隆子经酶切鉴定均有 0 .75kb片段的插入。测得编码 Sjc TPI的 DNA序列。结论 :用自行设计的引物成功地扩增了编码日本血吸虫大陆株 TPI抗原的基因片段并进行了克隆、测序 ,为制备重组 TPI进行疫苗试验打下基础。AIM:Cloning and sequencing the gene encoding TPI antigen from Chinese strain of S. japonicum.METHODS:RT- PCR technique was used to gain the target gene.After diges- tion with restriction enzyme,the gene was respectively cloned into the vectors M1 3mp1 8, M1 3mp1 9,then sequenced by the method of dideoxy- mediated chain- termination.RE- SUL TS:The 0 .75 kb c DNA encoding Sjc TPI antigen was amplified in vitro by RT- PCR us- ing total RNA as a template.0 .75 kb insert DNA was identified in six recombinantclones by restriction enzyme mapping. The DNA sequence of Sjc TPI was determined. CONCLUSION: The process have been made succesfully in the cloning and sequencing the gene encoding Sjc TPI.It will be scheduled to go ahead for vaccine trial probably using recombinant TPI antigen of S.japonicum.

关 键 词：日本血吸虫 大陆株 磷酸丙糖 异构酶 基因克隆 

分 类 号：R383.24[医药卫生—医学寄生虫学] Q78[医药卫生—基础医学]