免疫筛选恶性疟原虫cDNA克隆  

IMMUNOSCREENING OF c DNA CLONESEXPRESSING PLASMODIUM FALCIPARUM ANTIGEN BY USING DIFFERENT ANTIBODIES

作　　者：王燕妮[1] 谢毅[2] 肖谷田[2] 李明 毕惠祥 巢穗 王萍 李英杰 

机构地区：[1]贵州省遵义医学院免疫学教研室 [2]复旦大学遗传工程国家重点实验室,上海200433 [3]第一军医大学疟疾免疫研究室,广州510515

出　　处：《中国寄生虫学与寄生虫病杂志》1997年第4期193-197,共5页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：上海市青年科技启明星计划资助

摘　　要：目的 :获取恶性疟原虫 (海南株 )抗原的 c DNA克隆并进行初步鉴定。方法 :采用 dot-EL ISA,以兔免疫血清对恶性疟原虫 c DNA表达文库约 80万个重组噬斑进行筛选 ,并用 2 0株单克隆抗体和恶性疟患者血清对强阳性克隆进行再筛选。 PCR初步鉴定 17个强阳性克隆。结果 :兔免疫血清确定了 17个强阳性克隆 ,患者血清检测到 11个阳性克隆 (含 8个强阳性 ) ,11株单抗与 9个 c DNA克隆呈阳性反应。 17个强阳性克隆均能扩增出大小在 30 0 bp- 2 .5kb左右的条带。结论 :已筛选到能与抗恶性疟原虫兔血清、单克隆抗体及患者血清产生特异性免疫反应的 c DNA克隆。AIM:To screen c DNA genes of Plasmodium falciparum(FCC/HN) and to make pre- liminary identification.METHODS:8× 1 0 5 recombinants of Plasmodium falciparum c DNA library were screened using rabbit immune sera in dot- ELISA;the strong- positive clones were screened again using Mc Abs and sera from patients.Seventeenth strong- postive clones were preliminarily identitified by PCR.RESULTS:Fifty- eight positive clones including1 7 strong- positive ones were identified by immune sera of rabbits;eleven positive clones includ- ing8strong- positive ones were identified by sera of9patients.Nine clones were reacted with 1 1 strains of9Mc Abs.The inserts of1 7strong- positive clones were specifically amplified by PCR;their size ranged from ca.30 0 bp- 2 .5 kb.CONCLUSION:c DNA clones expressing F. falciparum antigen proteins were screened and would react with immune sera of rabbits, Mc Abs and sera of falciparum malaria patients.

关 键 词：恶性疟原虫 CDNA克隆 抗体 筛选 

分 类 号：R531.303[医药卫生—内科学] R382.31[医药卫生—临床医学]