[Construction of eukaryotic expression vector for HPC2 and its expression in HEK293 cells].

Kang-lian Tan, Zhi-jie Li, Jing-hua Liu, Hao Huang, Jing Tang, Peng Deng, Yong Jiang
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Abstract

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.

[HPC2真核表达载体的构建及其在HEK293细胞中的表达]。
目的:构建HPC2在HEK293细胞中表达的真核表达载体。方法:采用亚克隆法将pcDNA3/HPC2中的HPC2插入到标记载体pcDNA3-flag中。用常规脂质体法将重组质粒pcDNA3-flag/HPC2转染到HEK293细胞中。细胞裂解液用于Western blotting检测目标蛋白的表达。结果与结论:双限制性内切酶和DNA测序结果表明,HPC2真核表达载体构建成功,融合蛋白在HEK293细胞中高表达,为HPC2功能研究提供了重要依据。
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