日本血吸虫硫氧还蛋白谷胱甘肽还原酶基因克隆和序列分析  

Cloning and sequence analysis of thioredoxin glutathione reductase gene of Schistosoma japonicum

作　　者：谢曙英[1,2] 殷旭仁[3] 华万全[3] 曾小军[2] 陈红根[1,2] 梁幼生[3] 高琪[3] 余传信[3] 

机构地区：[1]南昌大学生命科学学院,南昌330000 [2]江西省寄生虫病防治研究所 [3]江苏省血吸虫病防治研究所,卫生部寄生虫病预防与控制技术重点实验室

出　　处：《中国血吸虫病防治杂志》2008年第6期427-430,共4页Chinese Journal of Schistosomiasis Control

基　　金：国家自然科学基金(30671833、30471515);江苏省医学重点人才基金(RC2007095);江苏省自然科学基金(BK2008110);江苏省科技厅公益专项基金(BM2007704)

摘　　要：目的克隆日本血吸虫硫氧还蛋白谷胱甘肽还原酶(SjTGR)基因,并进行序列分析。方法制备日本血吸虫成虫mRNA,反转录合成cDNA,采用聚合酶链反应(PCR)技术,从日本血吸虫成虫cDNA中扩增出SjTGR基因片段。通过TA克隆技术将该基因片段克隆到pGEM-T载体中进行DNA序列测定和生物信息学分析。结果采用PCR技术成功地从日本血吸虫成虫cDNA中扩增出TGR基因片段,TA克隆后DNA序列分析显示该基因长度为1791bp,编码596个氨基酸残基,理论分子质量65kDa,生物信息学分析显示与曼氏血吸虫TGR氨基酸序列同源性为82%,含有吡啶核苷酸二硫化物氧化活性中心CVNVGC序列,在氨基端含有谷氧还蛋白特征性的活性位点CPFC基序。结论SjTGR基因克隆获得成功,为发展抗日本血吸虫新药及疫苗候选分子奠定了基础。Objective To clone and analyze the gene of thioredoxin glutathione reductase of Schistosoma japonicum （Sj TGR）. Methods mRNA was extracted from an adult worm of S. japonicum for synthesizing cDNA by reverse transcriptase. The TGR gene amplified by PCR from the worms＇ eDNA was cloned into pGEM-T vector by TA clone technique. The gene was sequenced and its deduced protein structure was analyzed by the bioinformatics method. Results The gene fragment of Sj TGR was amplified by PCR and cloned into pGEM-T vector. The sequence data showed that Sj TGR gene was 1 791 bp in length, encoding a protein of 596 amino acids with a predicted of 65 kDa The homologous analysis showed 82% of identity between Sj TGR and TGR of S. mansoni. The Sj TGR contained the characteristic sequence of thiol-disulfide redox active motif of CVNVGC, and in addition, it also owned the active motif of CPFC similar to glutaredoxin in N terminal. Conclusions The gene of Sj TGR is cloned successfully and thus lays a foundation for developing new drugs and vaccine candidates against schistosome infection.

关 键 词：日本血吸虫 硫氧还蛋白谷胱甘肽还原酶 基因克隆 序列分析 

分 类 号：R383.24[医药卫生—医学寄生虫学]