兔抗鼠TGF-β1抗体对日本血吸虫尾蚴cDNA文库的免疫学筛选  

Immunology screening of schistosoma japonicum cercariae cDNA librarr with rabbit ANTI-MOUSE TGF-β1 antibody

作　　者：陈奕慧[1] 李孜[1] 李涛[1] 李巧艳[1] 麦璟莹[1] 

机构地区：[1]广州医学院病原生物学教研室,广东省广州市510182

出　　处：《中国现代医药杂志》2006年第12期58-61,共4页Modern Medicine Journal of China

摘　　要：目的为获得日本血吸虫(Schistosomajaponicum,Sj)转化生长因子β1(TGF-β1)基因的部分或全长cD-NA序列,同时验证用针对某一蛋白的抗体筛选表达文库是否可以得到相应蛋白对应的基因序列。方法采用兔抗鼠TGF-β1血清,对Sj尾蚴cDNA表达文库进行免疫学筛选,对阳性克隆进行PCR扩增后,将大于500bp的克隆进行复筛,对复筛阳性的克隆进行测序和生物信息学鉴定。结果对大约106个噬菌斑进行了初筛,共获得7个阳性克隆;经过PCR扩增后,其中有4个克隆大于500bp,复筛获得3个持续阳性克隆;对测序后的阳性克隆进行生物信息学鉴定,得到的3个克隆均与日本血吸虫辅酶Q10氧化还原酶(SjCHGC)基因具有高的同源性(Blastn分值都大于200)。结论SjCHGC蛋白与兔抗鼠TGF-β1抗体的反应为交叉反应,用抗某一蛋白的抗体筛选表达文库得到相应蛋白的基因序列的方法不一定可行。Objective To obtain the sequence of Schistosoma japonicum （Sj）transforming growth factorβ （TGF-β） gene, and to identify whether the antibody specific to one protein can be used to obtain the corresponding gene sequence by immunology screening cDNA expression library. Methods The cDNA library of Sj cercariae was screened with rabbit antimouse TGF-β1 antibody, and the inserts of positive clones were specifically amplified by PCR. Gene segments that exceeded 500bp were rescreened. The positive clones were sequenced. Then the sequences was compared with all sequences in Gen Bank database by Intemet. Results Of 106 phage plaques immunoscreened, 7 positive cDNA clones were found . Among them, four clones that exceeded 500bp were successfully amplified by PCR. Three continuing positive clones were confirmed. The sequencing data of 3 inserts by bioinformatics showed that they were high homologous to the Sj pankaryotic NADH-u- biquinone oxidoreductase （CHGC） in GenBank.（ Blastn score 〉200）. Conclusion It was cross reactivity between Sj CHGC protain and rabbit anti-mouse TGF-β1 antibody. The antibody specific to one protein probably can not be used to obtain the corresponding gene sequence by immunology screening cDNA expression library.

关 键 词：日本血吸虫 免疫学筛选 TGF-Β 

分 类 号：R392[医药卫生—免疫学]