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HPC2真核表达载体的构建及其在HEK293细胞中的表达 被引量：1

Construction of eukaryotic expression vector for HPC2 and its expression in HEK293 cells
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摘要 目的构建在哺乳动物细胞中表达的HPC2真核表达载体,并分析其在HEK293细胞中的表达。方法从重组pcDNA3/HPC2载体上将HPC2cDNA序列亚克隆至带有flag标记的真核表达载体pcDNA3-flag上,经PCR、酶切和测序鉴定,将重组质粒pcDNA3-flag/HPC2用脂质体瞬时转染HEK293细胞,细胞裂解后,用Westernblot分析并观察HPC2在HEK293细胞中的表达情况。结果PCR、酶切和DNA测序结果均表明重组质粒pcDNA3-flag/HPC2构建正确,并可在HEK293细胞内高效表达。结论成功构建了pcDNA3-flag/HPC2真核表达载体,该载体能在哺乳动物细胞中有效表达,为进一步研究HPC2的功能提供了重要的实验材料。 Objective To construct the eukaryotic expression vector for HPC2 for expression in HEK293 cells. Methods HPC2 from pcDNA3/HPC2 were inserted into the flag-tagged vector pcDNA3-flag by subcloning method. The recombinant plasmid pcDNA3-flag/HPC2 was then transfected into HEK293 cells using a routine lipofectamine method. The cell lysate was used for Western blotting to examine the expression of the target protein. Results and Conclusion Double restriction enzyme digestion and DNA sequencing indicated successful construction of the eukaryotic expression vector for HPC2 and the fusion protein was highly expressed in HEK293 cells, which provides an important basis for functional study of HPC2.
作者 谭康联 李志杰 刘靖华 黄浩 唐靖 邓鹏 姜勇
机构地区 南方医科大学广东省功能蛋白质组学重点实验室
出处 《第一军医大学学报》 CSCD 北大核心 2005年第12期1482-1484,1497,共4页 Journal of First Military Medical University
基金 国家重点基础研究发展项目(973)(2002CB513000) 国家高技术研究发展计划(863)(2001AA234061)~~
关键词 HPC2 载体构建 基因表达 转染 HPC2 vector construction gene expression transfection
分类号 Q78 [生物学—分子生物学]
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参考文献11

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第一军医大学学报
2005年 第12期

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