| 周洲，吴移谋，陈丽丽，李忠玉，陈超群.肺炎嗜衣原体CPAF原核表达载体的构建及其在大肠杆菌中的高效表达.[J].中南医学科学杂志.,2011,39(6):615-618. |
| 肺炎嗜衣原体CPAF原核表达载体的构建及其在大肠杆菌中的高效表达 |
| Construction of Prokaryotic Vector Encoding Chlamydophila Pneumoniae CPAF and Its Effective Expression in E.coli |
| 投稿时间：2010-11-26 |
| DOI： |
| 中文关键词: 肺炎嗜衣原体 CPAF基因 原核表达 |
| 英文关键词:Chlamydophila pneumoniae CPAF gene prokaryotic expression |
| 基金项目:国家自然科学基金（30901352）和2008湖南省教育厅研究生科研创新基金. |
|
| 摘要点击次数: 886 |
| 全文下载次数: 1067 |
| 中文摘要: |
| 目的对肺炎嗜衣原体蛋白酶样活性因子（CPAF）基因进行原核表达，为肺炎嗜衣原体的诊断及疫苗研究奠定基础。方法利用PCR技术从肺炎嗜衣原体标准株AR-39中扩增CPAF基因，将其克隆于原核表达载体pGEX-6p-2中，构建重组表达载体。并用双酶切、PCR及序列测定等方法进行鉴定后，在大肠杆菌中诱导表达GST融合蛋白。结果重组质粒中CPAF基因序列与肺炎嗜衣原体AR-39的同源性达到100%，CPAF在大肠杆菌中表达高效的GST融合蛋白。结论成功构建肺炎嗜衣原体CPAF基因的原核表达系统，并成功表达重组蛋白，为肺炎嗜衣原体疫苗的研究和临床检测试剂盒的研制打下基础。 |
| 英文摘要: |
| ObjectiveTo express the recombinant protein of chlamydial protease-like activity factor CPAF from Chlamydophila pneumoniae for using in diagnosis and vaccine development.MethodsThe CPAF gene was amplified from the genome of C.pneumoniae strain AR-39,cloned into prokaryotic expression vector pGEX-6p-2 for fusion expression with GST.With identification by sequencing,the recombinant plasmid was transformed into Escherichia coli XL1-Blue and the recombinant proteins fused with GST was analyzed by SDS-PAGE.ResultsSequence of the cloned CPAF gene was correct.The CPAF gene could be expressed in E.coli XL1-Blue which was transformed with pGEX-6p-2/CPAF but could not be expressed in E.coli XL1-Blue with pGEX-6p-2 or untransformed cells.ConclusionThe Prokaryotic expression plasmid pGEX-6p-2/CPAF was successfully constructed and expressed in E.coli XL1-Blue,which is useful in C.pneumoniae diagnosis and vaccine development. |
| 查看全文 查看/发表评论 下载PDF阅读器 |
| 关闭 |