论文部分内容阅读
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 ,计算机辅助设计和组合扩增技术是获得含多位点抗原决定簇的端粒酶催化亚基组合抗原融合蛋白的有效方法
OBJECTIVE: To use computer software to predict the telomerase catalytic subunit antigenic determinant, to prepare the recombinant telomerase catalytic subunit antigenic determinant, and to provide the conditions for preparing the monoclonal antibody of telomerase catalytic subunit. METHODS: The telomerase catalytic subunit epitopes were simulated by Goldkey software. A series of primers were designed to amplify the epitopes of the telomerase. The combined antigenic determinants were obtained by PCR amplification and cloned into pGEM_TEasyVector vector. The antigenic determinants The sequence was inserted into pGEX_6P_1 to induce the expression of the fusion protein, and the recombinant fusion protein was obtained by affinity chromatography. RESULTS: Four amino acid sequences of the most likely epitopes were found, and the recombinant human telomerase catalytic subunit fusion protein was obtained. CONCLUSIONS: The experimental results show that computer-aided design and combined amplification are effective methods to obtain the fusion protein of telomerase catalytic subunit with multi-site epitope