鼠反义转化生长因子βⅠ型受体真核表达质粒的构建与鉴定
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徐丽红, 郑 勇, 周 婷, 李 睿, 陈 莹
反义;质粒;转化生长因子βⅠ型受体;肝纤维化,徐丽红,郑勇,李睿,陈莹,周婷,徐丽红,通讯作者:,Constructionandidenti
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徐丽红, 郑勇, 李睿, 陈莹, 新疆石河子大学第一附属医院消化内科 新疆维吾尔族自治区石河子市 832008
周婷, 新疆石河子大学第一附属医院中心实验室 新疆省石河子市 832008
徐丽红, 女, 1970-08生, 汉族, 新疆石河子大学石河子医学院内科学在读硕士, 主治医师, 主要从事肝纤维化方面的研究.
兵团博士资金资助项目, No.05JC08
通讯作者: 郑勇, 832002, 新疆维吾尔族自治区石河子市, 新疆石河子大学第一附属医院消化内科. shzds-sh@xj.cninfo.net
收稿日期: 2005-10-25 接受日期: 2005-11-26
Construction and identification of rat pcDNA3.1(+)-antisense TβRI eukaryotic expressing plasmid
Li-Hong Xu, Yong Zheng, Ting Zhou, Rui Li, Ying Chen
Li-Hong Xu, Yong Zheng, Rui Li, Ying Chen, Department of Digestology, the First Affiliated Hospital of Shihezi University, Shihezi 832008, Xinjiang Uygur Autonomous Region, China
Ting Zhou, Center Laboratory, the First Affiliated Hospital of Shihezi University, Shihezi 832008, Xinjiang Uygur Autonomous Region, China
Supported by the Crops Foundation for the Doctor, No.05JC08
Correspondence to: Dr. Yong Zheng, Department of Digestology, the First Affiliated Hospital of Shihezi University, Shihezi 832008, Xinjiang Uygur Autonomous Region, China. shzds-sh@xj.cninfo.net
Received: 2005-10-25 Accepted: 2005-11-26
Abstract
AIM: To construct rat pcDNA3.1(+)-antisense TβRI (transforming growth factor beta receptor I) eukaryotic expressing plasmid, and to provide the experimental foundation for intervening the pathogenesis and development of liver fibrosis through TβRI.
METHODS: TRIzol assay was performed to obtain the total RNA from the rat liver tissues. The integrality, concentration and purity of total RNA were detected by ultraviolet spectrophotometry and agarose electrophoresis. The fragment of TβRI cDNA was obtained by reverse transcription polymerase chain reaction (RT-PCR), and then amplified by nest PCR. CaCl2 method was used to induce the susceptibility of cells. The eukaryotic expressing vector, at its multiple cloning sites, as well as the fragment of TβRI, was digested by EcoRⅠand Xhol, and then, was purified and retrieved by agarose electrophoresis separation ......
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