端粒酶催化亚基hTRT组合抗原决定簇基因的克隆与表达

Cloning and expression of combinatorial antigenic determinants of the catalytic subunit of human telomerase

目的 :利用计算机软件对端粒酶催化亚基抗原决定簇进行模拟预测分析 ,制备端粒酶催化亚基组合抗原决定簇重组蛋白 ,为制备端粒酶催化亚基单克隆抗体提供条件。方法 :以Goldkey软件模拟分析端粒酶催化亚基抗原决定簇 ,设计抗原决定簇的PCR扩增系列引物 ,组合PCR扩增获得组合抗原决定簇序列并克隆到pGEM_TEasyVector载体中 ,组合抗原决定簇基因序列插入pGEX_6P_1中 ,诱导表达融合蛋白 ,亲和层析获得组合融合蛋白重组抗原。结果 :发现了 4个可能性较大、序列较长的抗原决定簇氨基酸序列 ,获得了重组人端粒酶催化亚基组合融合蛋白。结论 :实验结果表明 。

Objective:To predict antigenic determinants of the catalytic subunit of human telomerase and to obtain recombinant protein of combinatorial antigenic determinants of the catalytic subunit of human telomerase. Methods: Antigenic determinants of the catalytic subunit of human telomerase was predicted and analyzed with Goldkey Program. Partial gene of antigenic determinants of the catalytic subunit of human telomerase was cloned and expressed in E.coli , then inserted in pGEX_6P_1 vector.Results:Six antigenic determinants were obtained by prediction, and recombinant protein of combinatorial antigenic determinants of the catalytic subunit of human telomerase was expressed.Conclusions:Computer aided analysis and combinatorial PCR are effective methods for obtaining the antigen protein of combinatorial antigenic determinants of the catalytic subunit of human telomerase.