期刊论文

中文

中国医药指南

1671-8194

2013

35

1-3

10.15912/j.cnki.gocm.2013.35.369

11A102:湖南省教育厅重点项目 12C0337:湖南省教育厅一般项目 2011KJ2:衡阳市科技局项目

本校为第一机构

目的：为了研制 Daxx-C 抗体并观察其在宫颈癌细胞中的应用效果，构建原核表达载体 pET28a/DM626-740，将其在大肠杆菌中进行诱导表达，纯化表达产物，鉴定其抗原性。方法用 PRIMER5.0软件设计 Daxx-C 基因片段（氨基酸626-740）特异性引物，引入BamHI 和 SalI 酶切位点，PCR 扩增目的基因。将 PCR 产物连入载体 pMD18-T，构建 pMD18-T/DM626-740；然后，将 BamHI 和 SalI 酶切产物连入载体 pET28a，经酶切鉴定及 DNA 序列分析后，构建原核表达载体 pET28a/DM626-740。将其转化 E.coli BL21，IPTG 诱导表达，利用 Ni 琼脂糖凝胶层析柱纯化表达产物，Western-blot 检测表达物与纯产物。结果双酶切...

Objective To prepare Daxx-C antibody and study its used effect on the cervical carcinoma, prokaryotic expression vector of Daxx-C626-740 was constructed and transformed into E.coli, and antigenicity of Daxx-C626-740 purified production was identified. Methods The specific primers of Daxx gene fragment(amino acids 626-740)including BamHI and SalI enzyme sites were esigned by software PRIMER5.0. PCR amplification products were inserted into MD18-T, and the vector of pMD18-T/DM626-740 was constructed. Then, DM626-740 gene fragments from pMD18-T/DM626-740 digested with BamHI and SalI were inserted...