神经药理学报

神经药理学报 ›› 2012, Vol. 2 ›› Issue (5): 19-23.

• 实验方法学 • 上一篇    下一篇

神经退行性疾病潜在致病肺炎衣原体蛋白Cpn0147酵母双杂交诱饵载体的构建

贾晓晖， 田颖新，田敏，韩小东，贾天军   

  1. 河北北方学院，病原生物学与免疫学研究所，张家口，075000，中国
  • 出版日期:2012-10-26 发布日期:2014-06-27
  • 通讯作者: 贾天军，男，教授，博士研究生，硕士生导师；研究方向：天然免疫与抗感染免疫；E-mail：jiantianjun@yahoo.com
  • 作者简介:贾晓晖，男，实验师，硕士研究生；研究方向：抗感染免疫；E-mail：jiaxiaohuistudent@163.com

Construction of Cpn0147 Bait Vector in Yeast Two-hybrid from Chlamydia Pneumonia Potentially Causing Neurodegenerative Diseases

JIA Xiao-hui, TIAN Ying-xin, TIAN Min, HAN Xiao-dong, JIA Tian-jun   

  1. Institute of Pathogen Biology and Immunology, Hebei North University, Zhangjiakou, 075000, China
  • Online:2012-10-26 Published:2014-06-27
  • Contact: 贾天军，男，教授，博士研究生，硕士生导师；研究方向：天然免疫与抗感染免疫；E-mail：jiantianjun@yahoo.com
  • About author:贾晓晖，男，实验师，硕士研究生；研究方向：抗感染免疫；E-mail：jiaxiaohuistudent@163.com

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摘要： 目的：构建Cpn0147酵母双杂交诱饵载体，为应用酵母双杂交系统筛选与其相互作用的蛋白奠定基础。方法： 应用PCR扩增技术获得Cpn0147基因片段，克隆入pGBKT7载体，鉴定构建是否成功，确定构建成功后将重组载体pGBKT7-Cpn0147转入AH109及Y187酵母中，进行诱饵载体的毒性及自激活活性分析。结果：Cpn0147诱饵载体构建成功，且该诱饵载体无自激活活性，对两种宿主酵母细胞无毒性。结论：诱饵载体pGBKT7-Cpn0147可用于酵母双杂交系统，为进一步筛选与之相互作用蛋白提供了实验基础。

关键词: 肺炎衣原体, 包涵体膜蛋白, 酵母双杂交, 诱饵载体, 自激活

Abstract: Objective: To construct a bait vector of pGBKT7-Cpn0147 for seeking its ligands with yeast two-hybrid system. Methods: Cpn0147 gene was amplified by polymerase chain reaction (PCR), and then cloned into pGBKT7 to construct the bait vector. These recombinant plasmids were transformed into yeast cells AH109 and Y187 to test toxic effects and self-activation activity. Results: The pGBKT7-Cpn0147 bait vector was constructed successfully, which neither had self-activation ability nor yeast cell toxicity. Conclusion: The constructed pGBKT7-Cpn0147 could be used as a bait vector in yeast two-hybrid system, and provided an experimental basis for further screening the interaction proteins.

Key words: chlamydia pneumoniae, inclusion, membrane protein, yeast , , two-hybrid system, bait , , vector, self-activation