水牛感染血清免疫筛选日本血吸虫成虫cDNA文库  

Immunoscreening of Schistosoma japonicum adult worm cDNA library by sera from infected Bubalus bubalis

作　　者：张冉[1] 易新元[2] 曾宪芳[2] 黄跃龙[2] Larry McReynolds 

机构地区：[1]湖南师范大学医学院检验系 [2]中南大学湘雅医学院寄生虫学教研室 [3]Molecular Parasitology Unit New England Biolabs,MA 01915,USA

出　　处：《中国地方病学杂志》2004年第6期549-552,共4页Chinese Jouranl of Endemiology

基　　金：WHO/TDR资助项目(980255)湖南省科委资助项目(00jzy2115)

摘　　要：目的 筛选新的有潜质的日本血吸虫保护性抗原分子编码基因。方法 用水牛感染血清对日本血吸虫(Schistosoma Japonicum,sj)成虫cDNA文库进行免疫筛选,将阳性克隆的插入片段进行PCR扩增和序列分析。结果 3轮筛选后,将7个阳性克隆经辅助噬菌体自动剪切并PCR扩增显示,插入的日本血吸虫cDNA片段大小在0.8-2.0 kb之间,选取其中4个经DNA测序分析,发现2个未曾报道过的日本血吸虫新基因,分别命名为sj-IB1和sj-Rho GTPase-like gene,并在Gene Bank登记注册。结论 水牛感染血清可识别日本血吸虫的特异性抗原分子,这些抗原分子的免疫保护作用值得进一步研究。Objective To explore the molecular mechanisms of the self-cure phenomenon of Bubalus bubalis (Bb) infected with Schistosomajaponicum (S. japonicum) and to find new vaccine candidates. Methods 5. japonicum adult worm cDNA library was screened with sera from infected Bb. The inserts of positive clones were amplified by PCR and the sequences were analyzed. Results Seven positive clones were obtained after rescreening. After automatically excised with helper phage, positive clones were further identified by PCR. The size of S. japonicum cDNA fragments in positive clones range from 0.8 to 2.0 kb. After DNA sequencing, it was discovered that there were two new S. japonicum genes of these clones. These new S. japonicum genes were designated Sj-IB1 and Sj-Rho GTP-like gene and registered in Gene Bank. Conclusions The serum of infected Bb could recognize S. japonicum specific antigens. It is worthwhile to further study immune protection of these cloned molecules.

关 键 词：水牛 免疫筛选 日本血吸虫 CDNA文库 感染血清 血吸虫病 

分 类 号：R346[医药卫生—基础医学]