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抑癌基因PTEN载体构建及在LOVO细胞中的表达 被引量：1

Vector construction of tumor suppressor gene PTEN and its expression in LOVO cells
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摘要 目的:构建抑癌基因PTEN的表达载体并在LOVO细胞中表达。方法:从人外周血中提取总RNA,通过RT-PCR扩增出PTEN基因片段,将PTEN基因连入pLenti6/V5载体。测序鉴定后,经脂质体转染入LOVO细胞,对细胞株进行RT-PCR和Western blot检测。结果:DNA测序证实pLenti6/V5-PTEN表达载体为阳性克隆;该表达载体转染LOVO细胞后上调了PTEN mRNA和蛋白的表达。结论:成功构建了pLen-ti6/V5-PTEN表达载体,为进一步研究PTEN在LOVO细胞中的功能奠定了基础。 Objective：To construct expression vector of PTEN gene and express the gene in LOVO cells.Methods： Extracted total RNA from human peripheral blood and amplified PTEN cDNA by RT-PCR,then the cDNA was inserted into vector pLenti6/V5.The positive clones were verified by sequencing and transfected to LOVO cells by lipofectamine.RT-PCR and Western blot were performed.Results： The pLenti6/V5-PTEN expression vector was constructed successfully after the confirmation by sequencing analysis.The pLenti6/V5-PTEN expression vector was transfected to LOVO cells,the mRNA and protein level of PTEN were upregulated obviously.Conclusion： The pLenti6/V5-PTEN was constructed successfully,and it provides a condition for the further investigation of PTEN function in LOVO cells.
作者 徐宁 李曼 黎小琼 赵学芹 何敏 黄宪章 庄俊华
机构地区 广州中医药大学第二附属医院
出处 《实验技术与管理》 CAS 北大核心 2011年第5期37-40,共4页 Experimental Technology and Management
基金 广东省自然科学基金项目(10151040701000031) 广东省中医院拔尖人才基金项目(201011-05)
关键词 PTEN 表达载体 LOVO细胞株 转染 PTEN expression vector LOVO cell line transfection
分类号 Q78 [生物学—分子生物学]
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参考文献9

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二级参考文献12

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共引文献10

同被引文献10

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实验技术与管理
2011年 第5期

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