{"title":"[Construction of eukaryotic expression vector for HPC2 and its expression in HEK293 cells].","authors":"Kang-lian Tan, Zhi-jie Li, Jing-hua Liu, Hao Huang, Jing Tang, Peng Deng, Yong Jiang","doi":"","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>To construct the eukaryotic expression vector for HPC2 for expression in HEK293 cells.</p><p><strong>Methods: </strong>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.</p><p><strong>Results and conclusion: </strong>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.</p>","PeriodicalId":11097,"journal":{"name":"Di 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2005-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Di 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
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.
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