日本血吸虫中国大陆株基因重组抗原Sj22.6kDa基因克隆与表达的研究  

STUDIESON THE CLONING AND EXPRESSION OF THE RECOMBINANT Sj2 2 .6k Da ANTIGEN GENE OF SCHISTOSOMA JAPONICUM

作　　者：张桂筠[1] 张兆松[2] 陈淑贞[2] 沈一平[2] 吴海玮[2] 苏川[2] 吴观陵[2] 

机构地区：[1]山西医科大学寄生虫学教研室,太原030001 [2]南京医科大学寄生虫学教研室,南京210029

出　　处：《中国寄生虫学与寄生虫病杂志》1997年第6期326-329,共4页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国务院总理专项研究基金!资助项目 ;编号 94- Y- 2 3

摘　　要：目的 :分离 Sj2 2 .6抗原基因 ,构建 Sj2 2 .6表达克隆。方法 :抗体筛选 Sj成虫 c DNA库 ,PCR扩增目的基因片段 ,亚克隆入表达载体 p GEX- 1λt。对重组体进行诱导表达并对表达产物进行 SDS- PAGE和 Western blot分析。结果 :PCR扩增产物为约 930 bp的 DNA条带。亚克隆入p GEX- 1λt,获得两个表达克隆 p GSj6和 p GSj2 4 ,特异性表达产物为 2 2 .6k Da抗原。重组体的表达方式为分离表达。结论 :从日本血吸虫成虫 c DNA库筛选到 Sj2 2 .6k Da克隆化基因 ,并构建了表达克隆 p GSj2 4和 p GSj6。AIM:To isolate Sj2 2 .6 gene and construct an expressive clone of r Sj2 2 .6 .METHODS: A S.japonicum adult c DNA library was screened by use of immune rabbit serum. From the possitive clones,one(λSj5 1 4,encoding Sj2 2 .6 ) was amplified by PCR to obtain its c DNA in- serts that was subcloned into the expressive plasmid vector p GEX- 1λtat Eco RI cloning site. The recombinants were induced by IPTG to express target protein which was analysed by SDS- PAGE and Western blotting. RESULTS:The specific productof PCR was about 930 bp which was subcloned into p GEX- 1 λt.Two positive expressive clones were obtained,p GSj6 and p GSj2 4,which expressed specific protein with a molecular weight of 2 2 .6 k Da. This re- combinant protein (r Sj2 2 .6 ) could be specifically recognized by immune rabbit serum.But the expressed product was not fusion protein,which meant that r Sj2 2 .6 was separated from the Sj GST that is encoded by the plasmid p GEX- 1 λt.CONCLUSION:Sj2 2 .6 antigen gene was screened from Sj adult c DNA library and subcloned into p GEX- 1λt,which resulted in two expressive clones,p GSj6 and p GSj2 4that expressed r Sj2 2 .6 .

关 键 词：日本血吸虫 基因克隆 重组抗原 

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