抗弓形虫单克隆抗体免疫筛选弓形虫速殖子cDNA文库

目的　探讨自制的抗弓形虫单克隆抗体 (McAb)相应抗原的分子结构机制 ,为弓形虫疫苗的研制寻找有效抗原提供依据。方法　用自制的抗弓形虫速殖子的McAb免疫筛选弓形虫速殖子cDNA文库 ,对阳性克隆cDNA插入片段进行PCR鉴定并测序分析。结果　 2个McAb第一轮分别获得 6个和 3个阳性克隆。分别进行第 2轮筛选 ,选取其中持续阳性最强的2个进行体外剪切 ,提取质粒DNA并进行PCR扩增和序列测定 ,经核苷酸同源性比较 ,可能为 2个新基因 ,其中 7C3-C3筛选的基因命名为WX2 (GenBank登录号为 :AY2 38892 ) ,2B9-G1筛选的基因命名为WX(GenBank登录号为 :AY2 0 8994 ) ,并用蛋白质分析软件对新基因编码的蛋白质结构和功能进行预测。结论　自制的具有一定保护性效果的McAb免疫筛选获得的阳性克隆插入基因片断的表达产物 ,可能具有抵抗弓形虫感染的作用 ,值得进一步深入研究。

Immuno-screening of the cDNA library of Toxoplasma gondii tachyzoite with monoclonal antibodies against Toxoplasma gondii

To explore the molecular mechanism of the corresponding antigen to the monoclonal antibody against Toxoplasma gondii in order to provide theoretic basis for the development of vaccines,the cDNA library of Toxoplasma gondii tachyzoites was immuno-screened with selfvaccines,the cDNA library of Toxoplasma gondii tachyzoites was immuno-screened with selfmade monoclonal antibodies (McAb),and the inserts of the positive clones were identified by PCR and sequencing analysis.In the first round,6 and 3 positive clones were obtained with two McAds respectively,and in the second round,selected two of the strongest persistently positive clones to be excised in vitro.The plasmid DVA was then extracted,amplified by PCR and undergone sequencing analysis.After comparative studies of nucleotide homology,these two genes might be the novel genes in which the gene screened with McAb 7C3-C was called WX2 (GenBank accession number AY238892),whereas that screened by McAb 2B9-G1 was called WX(GenBank accession number AY208994).Thus,two novel genes were obtained by means of immunoscreening with self-made monoclonal antibodies.Their molecular mechanism and immunological characterization wait for further investigation.