检索结果-南通市图书馆

Purification and characterization of biologically active recombinant human Eppin expressed in Escherichia coli

Chinese Medical Journal 2008年第7期121卷 620-624页

作者： ZHU Qing-yi GU Xiao-jian YANG Jin WANG Jun-hong TANG Bo WU Hong-fei Department ot Urology First Affiliated Hospital of Nanjing Medical University Nanjing Jiangsu 210029 China Department of Urology Jiangsu Province Hospital of TraditionalChinese Medicine Nanjing Jiangsu 210029 China Department of Biochemistry School of Life Science NanjingUniversity Nanjing Jiangsu 210093 China

Background Eppin （epididymis protease inhibitor） appears to play an important role in primate fertility. However, the function of Eppin and its antibody in men and its relationship with men＇s infertility are poorly studied. To reveal the significance and possibility of detection of anti-Eppin antibody in clinical infertilty cases, we developed an Escherichia coil expression system for the expression of biologically active human Eppin. Methods The human Eppin gene was cloned into PET-28a（＋） vector after induction with 0.5 mmol/L isopropy-β-Dthiogalactoside （IPTG） at 26℃ for 4 hours, and the expressed fusion protein His6-Eppin was purified by Ni2. affinity chromatography. Afterwards, six female 8-week-old Balb/c mice were immunized with purified His6-Eppin for three weeks. Their sera were collected and polyclonal antibodies against His6-Eppin were purified, all of which were further verified by Western-blot and immunofluorescence analysis. Results About 18.33 mg His6-Eppin was obtained from 1-L flask culture. The produced polyclonal antibodies against His6-Eppin recognized the Eppin protein both in human epididymis and in HEK293T cells by over-expression of the recombinant human Eppin. Conclusion The purified His6-Eppin protein has biological activity, which might be a candidate for clinical diagnosis of infertility and development of male immuno-contraceptive agents.

关键词： epididymis protease inhibitor expression purification biological function

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

欢迎您,

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

请选择保存的检索档案：

请选择收藏分类：

通借通还

欢迎您,

建议与咨询 留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

请选择保存的检索档案： 新增检索档案 确定 取消

请选择收藏分类： 新增自定义分类 确定 取消

通借通还

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

请选择保存的检索档案：

请选择收藏分类：