| TSLC1真核表达载体的构建及在HepG2肝癌细胞中的表达 |
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| 引用本文： | 肖钟迪,张玉成,房学东. TSLC1真核表达载体的构建及在HepG2肝癌细胞中的表达[J]. 中国体视学与图像分析, 2010, 0(1): 89-94 |
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| 作者姓名： | 肖钟迪 张玉成 房学东 |
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| 作者单位： | 吉林大学第二临床医院;大庆油田总医院普外科;吉林大学中日联谊医院中心研究室; |
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| 基金项目： | 吉林省发展与改革委员会资助课题[(2006)1550号] |
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| 摘 要： | 目的构建TSLC1真核表达载体,并转染到肝癌细胞HepG2中使之表达,为研究TSLC1的抗肝癌作用奠定基础。方法利用RT-PCR技术扩增TSLC1基因全长,并与真核表达载体pRe-ceiver-M29进行连接;应用双酶切、PCR以及测序鉴定此连接载体;脂质体Lipofectamine 2000介导重组载体转染到HepG2细胞中,经G418筛选建立稳定转染细胞株,分别采用RT-PCR和免疫组化法检测其表达。结果RT-PCR获得了约1 329 bp大小的TSLC1基因片段;经过双酶切、PCR以及测序鉴定证实TSLC1基因片段正确插入真核表达载体pReceiver-M29中;与对照组比较,RT-PCR方法可见显示转染组细胞TSLC1mRNA表达明显增多,免疫组化法可见显示转染组细胞TSLC1蛋白表达明显增高。结论成功构建了真核表达载体pReceiver-M29-TSLC1,建立了稳定转染TSLC1的HepG2细胞株。
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| 关 键 词： | TSLC1 聚合酶链反应 克隆 HepG2 |
Construction of an eukaryotic expressing vector pReceiver-M29-TSLC1 and its expression in hepatoma HepG2 cells |
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| XIAO Zhongdi,,ZHANG Yucheng,FANG Xuedong. Construction of an eukaryotic expressing vector pReceiver-M29-TSLC1 and its expression in hepatoma HepG2 cells[J]. Chinese Journal of Stereology and Image Analysis, 2010, 0(1): 89-94 |
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| Authors: | XIAO Zhongdi ZHANG Yucheng FANG Xuedong |
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| Affiliation: | XIAO Zhongdi1,2,ZHANG Yucheng3,FANG Xuedong1(1.The Second Hospital of Jilin University,Changchun 130041,China,2.Department of General Surgery,Daqing Oil Field General Hospital,Daqing 163001,3.Central Laboratory,China-Japan Union Hospital,Jilin University,Changchun 130033,China) |
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| Abstract: | Objective To construct an eukaryotic expressing vector pReceiver-M29-TSLC1 and examine its expression in hepatoma HepG2 cells.Methods The full length of TSLC1 was amplified and the pReceiver-M29-TSLC1 was constructed by polymerase chain reaction(PCR).The pReceiver-M29-TSLC1 was identification using enzyme cutting,PCR and sequencing.After the pReceiver-M29-TSLC1 was transfected into hepatoma HepG2 cell mediated with Lipofectamine 2000,a stable transfected cell line was established using G418 screening and th... |
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| Keywords: | TSLC1 PCR clone hepatoma cell line HepG2 |
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