日本血吸虫尾蚴抗原免疫血清筛选成虫cDNA文库与新基因分析  

Immunoscreening of Schistosoma japonicum Adult Worm cDNA Library with Sera Vaccinated with Cercaria Antigen and Analysis of Novel Genes

作　　者：袁仕善[1] 易新元[1] 曾宪芳[1] 欧阳理[1] 王敏[1] 唐连飞[1] Larry Mc Reynolds 

机构地区：[1]中南大学湘雅医学院血吸虫病研究室 [2]Molecular Parasitology Group,New England Biolabs,MA01915,USA

出　　处：《中国寄生虫学与寄生虫病杂志》2003年第5期289-292,共4页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：湖南省科技厅资助项目 (No.0 0jzy2 1 1 5);WHO/TDR资助项目 (No.980 2 68)

摘　　要：目的　筛选与日本血吸虫尾蚴抗原有共同免疫原性的日本血吸虫成虫抗原分子 ,为血吸虫病诊断和疫苗研究提供新的候选分子。　方法　利用日本血吸虫尾蚴可溶性抗原免疫兔血清筛选日本血吸虫 (Schistosomajapon icum ,Sj)成虫cDNA文库 ,对阳性克隆的插入片段进行PCR扩增和序列分析。　 结果　共筛选出 13个阳性克隆 ,PCR扩增出特异的插入片段。测序分析获 4个新基因SjCAI、SjCA、SjCAI 2和SjCAI 3 (登录号分别为AF495 883、AF5 15 83 4、AY1180 86和AY12 93 0 3 ) ,分别编码 3 5 3、161、13 7和 72个氨基酸的蛋白。SjCAI蛋白含 6个DNA结合锌指 ,与原肠胚锌指蛋白XLCGF48.2有一定同源性 ;SjCA蛋白、SjCAI 2蛋白和SjCAI 3蛋白含N 糖基化和磷酸化位点。　 结论　筛选Sj成虫cDNA文库获得了新的基因序列。Objective To explore antigens possessing common immunogenicity with Schistosoma japonicum (Sj) cercariae antigens, and to find out new candidate antigens for schistosomiasis diagnosis and vaccine. Methods Sj adult cDNA library was screened using sera from rabbits vaccinated with Sj cercariae antigen, the inserts of positive clones were amplified by PCR, all positive clones were sequenced and the data were analysed using Nucleotide BLAST software of NCBI and Expert Protein Analysis System of Swiss Institute of Bioinformatics. Results Thirteen positive clones were obtained after three rounds of immunoscreening, and all amplified by PCR.Among four novel genes, SjCAI, SjCA, SjCAI-2 and SjCAI-3(GenBank accession number:AF495883, AF515834, AY118086 and AY129303,respectively)encoded proteins with 353, 161, 137 and 72 animo acids respectively. Sj CAI protein contained six DNA-binding zinc fingers and showed some homology to gastrula zinc finger protein XLCGF48.2; proteins encoded by SjCA, SjCAI-2 and SjCAI-3 respectively contained N-glycosylation sites and phosphorylation sites. Conclusion Novel genes were obtained by immunoscreening Sj adult cDNA library.

关 键 词：日本血吸虫 尾蚴 抗原 免疫血清筛选 成虫 CDNA文库 新基因分析 

分 类 号：R383.24[医药卫生—医学寄生虫学]