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[用尾蚴抗原免疫血清筛选日本血吸虫成虫cDNA文库及新基因分析]

[Immunoscreening of Schistosoma japonicum adult worm cDNA library with sera vaccinated with cercaria antigen and analysis of novel genes].

作者信息

Yuan Shi-shan, Yi Xin-yuan, Zeng Xian-fang, Ouyang Li, Wang Min, Tang Lian-fei, McReynolds Larry

机构信息

Laboratory of Schistosomiasis, Xiangya School of Medicine, Central South University, Changsha 410078.

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2003;21(5):289-92.

Abstract

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.

摘要

目的

探索与日本血吸虫尾蚴抗原具有共同免疫原性的抗原,寻找血吸虫病诊断和疫苗的新候选抗原。

方法

用日本血吸虫尾蚴抗原免疫兔血清筛选日本血吸虫成虫cDNA文库,通过PCR扩增阳性克隆的插入片段,对所有阳性克隆进行测序,并使用NCBI的核苷酸BLAST软件和瑞士生物信息学研究所的专家蛋白质分析系统对数据进行分析。

结果

经过三轮免疫筛选获得13个阳性克隆,均经PCR扩增。在4个新基因中,SjCAI、SjCA、SjCAI-2和SjCAI-3(GenBank登录号分别为:AF495883、AF515834、AY118086和AY129303)分别编码含353、161、137和72个氨基酸的蛋白质。Sj CAI蛋白含有6个DNA结合锌指,与原肠胚锌指蛋白XLCGF48.2有一定同源性;SjCA、SjCAI-2和SjCAI-3分别编码的蛋白含有N-糖基化位点和磷酸化位点。

结论

通过免疫筛选日本血吸虫成虫cDNA文库获得了新基因。

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