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p16基因真核表达质粒的构建及其在骨肉瘤细胞中的表达 被引量：1

Construction of eukaryotic expression vector for p16 gene and its expression in osteosarcoma cells
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摘要 目的构建p16基因真核表达质粒,并鉴定其在骨肉瘤U-2OS细胞中的表达。方法提取HeLa细胞总RNA,扩增p16基因,克隆至真核表达质粒pcDNA3-HA中,构建重组表达质粒pcDNA3-HA-p16,转染U-2OS细胞,免疫荧光及Westernblot法鉴定p16/HA的表达。结果重组表达质粒pcDNA3-HA-p16经双酶切及测序证实构建正确;p16/HA在U-2OS细胞中成功表达,且主要分布于细胞核。结论成功构建了p16基因真核表达质粒,并在U-2OS细胞中成功表达。 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-pl6 was transfected to U-2OS cells, and the expression of pl6/HA was identified by IFA and Western blot. Results Restriction analysis and sequencing proved that recombinant plasmid pcDNA3-HA-pl6 was constructed correctly. The p16/HA was successfully expressed in U-20S cells, which was mainly distributed in nucleus. Conclusion The eukaryotic expression vector for p16 gene was successfully constructed and expressed in U- 2OS cells.
作者 夏羿凡 余娴 张健
机构地区 重庆医科大学附属第一医院骨科 川北医学院附属医院骨科 川北医学院药物研究所
出处 《中国生物制品学杂志》 CAS CSCD 2012年第2期190-192,共3页 Chinese Journal of Biologicals
关键词 基因 P16 真核细胞 基因表达 骨肉瘤 Gene, p16 Eukaryotic cells Gene expression Osteosarcoma
分类号 Q78 [生物学—分子生物学] R738.1 [医药卫生—肿瘤]
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参考文献15

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同被引文献10

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中国生物制品学杂志
2012年 第2期

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