期刊论文

Constructionof BTF Red Fluorescent Protein Expression Plasmid and Its Expression in Rat VSMCs

中文

医学分子生物学杂志

1672-8009

2014

3

161-164

10.3870/j.issna.1672-8009.2014.03.007

(81200881):国家自然科学基金青年项目 (No．12JJ6073):湖南省自然科学基金 (81200881):This work was supported by grants from the National Natural Science Foundation of China (12JJ6073):the Natural Science Foundation of Hunan Province

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目的：构建Bcl-2相关转录因子1（BTF）与红色荧光蛋白（RFP）融合表达载体并在大鼠血管平滑肌VSMC细胞中表达，为进一步研究BTF的相互作用蛋白及作用机制奠定实验基础。方法以人体外周血白细胞cDNA为模板， PCR扩增Btf基因，插入红色荧光蛋白表达载体pDsRed-N1中，构建重组表达载体pDsRed-N1-Btf，转染大鼠血管平滑肌VSMC细胞，通过激光共聚胶显微镜观察其在转染细胞中的表达及定位。结果 DNA测序、酶切鉴定的结果显示，红色荧光蛋白表达载体pDsRed-N1-Btf构建成功，且序列正确。转染后经激光共聚胶显微镜观察到VSMC细胞中有红色荧光，且BTF主要定位于细胞质中。结论成功构建了pDsRed-N1-Btf表达...

Objective To construct the expression vector of Bcl-2-associated transcription fac-tor 1 (BTF) fused to red fluorescent protein ( RFP) and examine its expression in rat vascular smooth muscle cells ( VSMCs) in order to provide an experimental foundation for further studying the interaction of BTF and other proteins in cells and the action mechanism .Methods Btf gene was amplified by PCR with the cDNA of peripheral white blood cells as the template , and inserted into the red fluorescent protein expression vector pDsRed-N1 to obtain the recombinant plasmid pDsRed-N1-Btf, which was then transfec...