摘要
目的：获取编码可能指示再感染倾向的日本血吸虫特异免疫分子的ｃＤＮＡ克隆。方法：用日本血吸虫病流行区化疗后再感染人群高特异性ＩｇＧ４水平的混合血清筛选构建于表达载体λｇｔ１１的日本血吸虫成虫ｃＤＮＡ文库，以ＰＣＲ鉴定免疫筛选的阳性ｃＤＮＡ克隆插入片段选择插入片段大的克隆进行序列测定及基因数据库同源性检索。结果与结论：对ｃＤＮＡ文库中约８．６×１０４个噬菌斑进行了筛选，有１１个噬菌斑呈阳性反应，其中有可能编码日本血吸虫的线粒体蛋白的基因克隆及可能与人群日本血吸虫再感染相关的基因克隆等４个日本血吸虫基因克隆。
AIM:To obtain cDNA clones coding for S. japonicum(Sj) reinfection related specific immunologic molecules for vaccination or diagnosis. METHODS: An adult S.japonicum cDNA library, of which the cloning efficiency is 3.13×10 6, with directional inserts in the vector lambda gt11, was screened with pooled serum from 43 reinfected human cases in a schistosomiasis endemic area with high level of Sj specific lgG4 antibodies. Primers located several bases away from the insert position on the lambda gt11 arm were used to amplify the insert fragments by standard PCR. SILVER SEQUENCE TM (PCR silver) DNA sequencing system was then used to sequence selected positive clones with relatively large inserts and homology comparison with GenBank database was also carried out. RESULTS AND CONCLUSION: Eleven phage plaques were defined as positive out of the 8.6×10 4 phage plaques immunoscreened by human specific lgG4 antibody. GenBank database retrieval showed that 4 new Sj gene clones including Sj reinfection related specific protein encoding gene and Sj mitochondrial protein coding gene have been obtained.
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
1998年第5期372-375,共4页
Chinese Journal of Parasitology and Parasitic Diseases
基金
国家自然科学基金
总理预备金所设专项基金
