一个广州管圆线虫特异性抗原候选基因的克隆与表达  

Cloning and expressing of a specific antigen cDNA candidate of Angiostrongylus cantonensis

作　　者：孟锦绣[1] 詹希美[2] 程梅[2] 李素丽[2] 甘明[2] 徐贵峰[2] 李卓雅[2] 何蔼[2] 

机构地区：[1]广东省人民医院医学研究中心,广州510080 [2]中山大学中山医学院,广州510089

出　　处：《中国人兽共患病学报》2006年第8期704-707,711,共5页Chinese Journal of Zoonoses

基　　金：国家"十五"科技攻关重大项目(2003BA712A03-07);国家自然科学基金(301708377);广州市科技三项(2005J3-7561);广东省人民医院科学技术研究基金(Y200522)资助

摘　　要：目的用免疫探针交叉筛选前期实验获得的广州管圆线虫候选抗原基因,将筛选出的特异抗原候选基因进行原核表达、鉴定。方法制备抗血吸虫和抗旋毛虫多克隆抗体血清,交叉筛选广州管圆线虫抗原候选基因,从中获得特异性抗原候选基因,将其从重组噬菌体状态转化为质粒状态,再亚克隆入原核表达载体pET-30a-c(+)进行融合表达,SDS-PAGE鉴定表达产物,同时对该基因进行生物信息学分析。结果获得了一个广州管圆线虫特异性抗原候选基因,经生物信息学分析该抗原基因属中间纤维家族基因(intermediate filament,IF),原核表达产物为48KDa。结论获得了一个广州管圆线虫特异性抗原候选基因及其原核表达产物。In order to specific candidate antigen of Angiostrongylus cantonensis with cross-immunoscreening the cDNA, and subcloning, expressing and identifying the specifi antigen. The antigens, previously obtained by screening the A. cantonensis cDNA library by anti-serum, were tested by cross-reaction with other parasite anti-serum to get specific antigen candidate cDNA. The recombinant phages inserted the specific antigen cDNA segment were transformed into plasmids. The specific antigen cDNA were subcloned into pET-30a-c（＋） expression vectors and expressed in vitro. The expressed products were identified by SDS-PAGE. Bioinformatics were used to analyse the cDNA sequence. The results showed a specific antigen cDNA of A. cantonensis was obtained, the cDNA sequence belongs to intermediate filament （IF） family by bioinformatics analyses. The expression product is 48kDa. It is evident that and A specific antigen cDNA and its expression products were obtained.

关 键 词：广州管圆线虫 抗原 中间纤维 CDNA文库 

分 类 号：R532.1[医药卫生—内科学] R737.250.4[医药卫生—临床医学]