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.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
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
关键词
广州管圆线虫
抗原
中间纤维
CDNA文库
Angiostrongylus cantonensis, antigen, intermediate filament, cDNA library
