| 抑癌基因PTEN载体构建及在LOVO细胞中的表达 |
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| 引用本文： | 徐宁,李曼,黎小琼,赵学芹,何敏,黄宪章,庄俊华.抑癌基因PTEN载体构建及在LOVO细胞中的表达[J].实验技术与管理,2011,28(5):37-40. |
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| 作者姓名： | 徐宁 李曼 黎小琼 赵学芹 何敏 黄宪章 庄俊华 |
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| 作者单位： | 广州中医药大学第二附属医院,广东广州,510120 |
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| 基金项目： | 广东省自然科学基金，广东省中医院拔尖人才基金 |
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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。
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| 关 键 词： | PTEN 表达载体 LOVO细胞株 转染 |
Vector construction of tumor suppressor gene PTEN and its expression in LOVO cells |
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| Xu Ning,Li Man,Li Xiaoqiong,Zhao Xueqin,He Min,Huang Xianzhang,Zhuang Junhua.Vector construction of tumor suppressor gene PTEN and its expression in LOVO cells[J].Experimental Technology and Management,2011,28(5):37-40. |
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| Authors: | Xu Ning Li Man Li Xiaoqiong Zhao Xueqin He Min Huang Xianzhang Zhuang Junhua |
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| Institution: | (Second Affiliated Hospital,Guangzhou University of Chinese Medicine,Guangzhou 510120,China) |
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| Abstract: | 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. |
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| Keywords: | PTEN expression vector LOVO cell line transfection |
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