2012•Zhongguo shengwuzhipinxue zazhiRequires access
Construction of eukaryotic expression vector for p16 gene and its expression in osteosarcoma cells
Zhang Jian
Open publisher page 0 citations
Abstract
Objective To construct a eukaryotic expression vector for p16 gene and determine its expression in osteosarcoma U-2OS cells.Methods Total RNA of HeLa cells was extracted,with which p16 gene was amplified and cloned into eukaryotic expression vector pcDNA3-HA.The constructed recombinant plasmid pcDNA3-HA-p16 was transfected to U-2OS cells,and the expression of p16/HA was identified by IFA and Western blot.Results Restriction analysis and sequencing proved that recombinant plasmid pcDNA3-HA-p16 was constructed correctly.The p16/HA was successfully expressed in U-2OS cells,which was mainly distributed in nucleus.Conclusion The eukaryotic expression vector for p16 gene was successfully constructed and expressed in U-2OS cells.