Construction of eukaryotic expression of CIB vectors and its expression in NIH3T3 cells

HUI Ling; WANG Xiao-hui; YUAN Bi-bo; YANG Jin-kai; LU Tong-de; HUI Bin; CAI Wen-qin

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Construction of eukaryotic expression of CIB vectors and its expression in NIH3T3 cells

更新时间：2025-01-15

- Construction of eukaryotic expression of CIB vectors and its expression in NIH3T3 cells
- Journal of Army Medical University Vol. 29, Issue 3, Pages: 216-218(2007)
- 作者机构：
  
  1. 兰州军区兰州总医院医学实验中心
  2. 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
  3. 复旦大学上海医学院药理研究中心
- 作者简介：
- 基金信息：
- DOI：
  CLC： Q513;Q782;Q786
- Published：2007，
  
  Published Online：15 February 2007，
- 稿件说明：
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HUI LING, WANG XIAO-HUI, YUAN BI-BO, et al. Construction of eukaryotic expression of CIB vectors and its expression in NIH3T3 cells. [J]. Journal of army medical university, 2007, 29(3): 216-218.
DOI：

HUI LING, WANG XIAO-HUI, YUAN BI-BO, et al. Construction of eukaryotic expression of CIB vectors and its expression in NIH3T3 cells. [J]. Journal of army medical university, 2007, 29(3): 216-218. DOI：

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.

关键词

CIB基因克隆真核表达

Keywords

calcium- and integrin-binding proteingene cloningeukaryotic expression

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