中国医科大学学报

中国医科大学学报

中国医科大学学报 ›› 2013, Vol. 42 ›› Issue (11): 970–973.

• 论著 • 上一篇    下一篇

Cav1.2钙通道片段CT1融合蛋白的制备及生物学活性鉴定

封瑞1,胡慧媛1,郭凤1,于丽凤1,赵美眯1,龟山正树2,郝丽英1   

  1. 1中国医科大学药学院药物毒理学教研室,沈阳110001;
    2鹿儿岛大学医学部,鹿儿岛890-8544,日本
  • 收稿日期:2013-09-11 出版日期:2013-11-30 发布日期:2013-11-28
  • 作者简介:封瑞(1981-),女,讲师,硕士.
  • 基金资助:
    国家自然科学基金(31071004

Purification of CT1 fragment of Cav1.2 channel and Identification of Its Biologic Activity

FENG Rui1,HU Hui-yuan1,Guo Feng1,Yu Li-feng1,Zhao Mei-mi1,M Kameyama2,HAO Li-ying1   

  1. 1.Department of Pharmaceutical Toxicology,School of Pharmacy,China Medical University,Shenyang 110001,China;
    2.Department of Physiology,Graduate School of Medical & Dental Sciences,Kagoshima University,Kagoshima 890-8544,Japan
  • Received:2013-09-11 Online:2013-11-30 Published:2013-11-28

摘要: 目的诱导表达Cav1.2钙通道片段CT1与谷胱甘肽转移酶(GST)重组的融合蛋白并进行纯化和鉴定。方法将pGEX-6p-3/CT1重组质粒转化入E.coliBL21中并诱导表达,采用非离子去污剂B-per的方法分离纯化GST-CT1融合蛋白,使用Westernblot技术鉴定GST-CT1融合蛋白,pull-downassay实验方法检测GST-CT1融合蛋白的生物学功能。结果非离子去污剂B-per能够成功分离纯化GST-CT1融合蛋白,识别此片段的特异性抗体能够检测到GST-CT1融合蛋白;制备获得的GST-CT1融合蛋白具有与钙调蛋白结合的生物学功能。结论技术简单、操作快速的非离子去污剂B-per能够分离纯化具有生物学功能的GST-CT1融合蛋白。

关键词: 融合蛋白, 非离子去污剂, 钙通道, 片段

Abstract: Objective To purify the CT1 fragment of Cav1.2 channel and investigate its biologic activity. Methods Escherichia coliBL21 was transformed with plasmid of pGEX- 6p-3/ CT1. The transformed Escherichia coliBL21 were induced by isopropyl-#x003b2;-D-thiogalactoside (IPTG). Then,the GST-CT1 fusion protein was purified by nonionic detergent B-per method,and identified by Western Blot. In addition,the pull-down assay was performed to investigate the

Key words: fusion protein, nonionic detergent, calcium channel, fragment

中图分类号: 

  • R965
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