[1]惠玲,王晓辉,袁碧波,等.钙整合素结合蛋白CIB真核表达载体的构建及在NIH3T3细胞中的表达[J].陆军军医大学学报(原第三军医大学学报),2007,29(03):216-218.
 HUI Ling,WANG Xiao-hui,YUAN Bi-bo,et al.Construction of eukaryotic expression of CIB vectors and its expression in NIH3T3 cells[J].J Amry Med Univ (J Third Mil Med Univ),2007,29(03):216-218.
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钙整合素结合蛋白CIB真核表达载体的构建及在NIH3T3细胞中的表达(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
29卷
期数:
2007年第03期
页码:
216-218
栏目:
论著
出版日期:
2007-02-15

文章信息/Info

Title:
Construction of eukaryotic expression of CIB vectors and its expression in NIH3T3 cells
作者:
惠玲王晓辉袁碧波杨金开吕同德惠斌蔡文琴
兰州军区兰州总医院医学实验中心;第三军医大学基础医学部神经生物学教研室,重庆市神经科学研究所;复旦大学上海医学院药理研究中心
Author(s):
HUI Ling WANG Xiao-huiYUAN Bi-bo YANG Jin-kai LU Tong-de HUI Bin CAI Wen-qin
Experimental Center of Medicine, Lanzhou General Hospital of Lanzhou Command, Lanzhou 730050;Department of Histology and Embryology, Third Military Medical University, Chongqing 400038;Pharmacology Research Center of Shanghai Medical College, Fudan University, Shanghai 200032, China
关键词:
CIB基因克隆真核表达
Keywords:
calcium- and integrin-binding protein gene cloning eukaryotic expression
分类号:
Q513;Q782;Q786
文献标志码:
A
摘要:
目的     构建钙整合素结合蛋白(calcium- and integrin-binding protein, CIB)的真核表达载体,了解它在真核细胞中的表达及定位。     方法     从人脑组织分离总RNA,采用RT-PCR获得CIB开放读框序列,克隆至真核表达载体pCMV-myc,构建表达带myc表达标签的真核表达载体pCMV-myc-CIB,将其转染至NIH3T3细胞中,免疫组化鉴定其在细胞中的表达及定位。     结果     序列分析表明克隆的CIB序列与GeneBank报道的序列一致,脂质体转染入NIH3T3细胞中,用myc抗体检测到目的蛋白myc-CIB的表达。     结论     pCMV-myc-CIB表达载体构建成功,并在真核细胞中得到了正确表达,CIB蛋白主要定位于细胞质中。
Abstract:
Objective     To construct the expression vector of calcium- and integrin-binding protein (CIB), and study its localization as a fusion protein in NIH3T3 cell.      Methods     Total RNA was extracted from human brain tissue for CIB sequence with RT-PCR, and then the obtained fragment was ligated into pCMV-myc vector. After confirmation with sequence analysis, the expression vector pCMV-myc-CIB was transformed into the NIH3T3 cell by Lipofectamin 2000. The protein expression was observed by immunofluorescent staining.      Results     DNA sequence analysis showed that the CIB cDNA had been successfully inserted into pCMV-myc expression vector. myc-CIB was expressed in the NIH3T3 cells transfected with pCMV-myc-CIB.      Conclusion     The eukaryotic expression vector containing CIB is successfully constructed and expressed in the cytoplasma of NIH3T3 cells.

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更新日期/Last Update: 2008-11-13