Expression of Recom binant pEGFP·HPV16 E7 in Eukaryocyte

ZUO Ze—hua．LI Hui．WU Xin—xing

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August 2003

ZUO Ze—hua．LI Hui．WU Xin—xing, Expression of Recom binant pEGFP·HPV16 E7 in Eukaryocyte[J]. Virologica Sinica, 2003, 18(4): 344-347.

Citation: ZUO Ze—hua．LI Hui．WU Xin—xing, . Expression of Recom binant pEGFP·HPV16 E7 in Eukaryocyte .VIROLOGICA SINICA, 2003, 18(4) : 344-347.

pEGFP—HPV16 E7重组融合蛋白质粒的构建及其表达

1.
武汉大学医学院病毒学研究所分子生物学研究室，湖北武汉，430071

摘要

摘要：应用基因重组技术，构建增强绿色荧光蛋l~(EGFP)与人乳头瘤病毒16型E7(HPV16E7)的重组融合表达质粒，经限制性内切酶酶切鉴定和PCR 分析后，用基因转染技术将其导入小鼠肝癌细胞，荧光显微镜下观察融合蛋白的表达。酶切鉴定和PCR 分析证实重组质粒中插入目的基因片段的大小、方向和插入位点均正确，在转染的小鼠肝癌细胞中观察到绿色荧光蛋白的表达。构建的pEGFP．HPV16E7融合表达质粒能直观地反映转染细胞中EGFP-HPV16E7融合蛋白的表达。由于转化率与表达率融为一体，故有利于对转染细胞的筛选，缩短转染细胞在体外的筛选的时间适用于对HPV16E7 分子生物学特性、致瘤机理及APC 提呈等的研究。为建立表达HPV16E7的实体瘤动物模型奠定了基础。
- 人乳头瘤病毒
- , 表达载体
- , 基因转染
- , 真核表达

Expression of Recom binant pEGFP·HPV16 E7 in Eukaryocyte

1.
Virus Research Institution ofMedical College，Wuhan University Wuhan 430071，China

Abstract

Abstract：In order to make bases for construction of animal tumor model which expresses EGFP— HPV l 6E7，recombinant expression plasmid was constructed by techniques of gene recombination after screening an d identifing by restriction and PCR．Then the recombinant was tran sfected into mouse liver cancer cell by techniques of gene transfection and detected expression by fluoroscopy．pEGFP—HPV 1 6 E7 was identified by enzyme digestion an d PCR．Th e resent showed that the length ．inserted location and direction of the target gene was correct an d the expression of EGFP in tran sfected cell was observed Because of it’S pEGFP—HPV 1 6E7 expression plasmid that mak es it easy to assess the expression of EGFP-HPV 1 6E7 fusion protein and to sift the transfected cells．
- Human papillomavirus
- , Eukaryotic expression
- , Expression vector
- , e7 gene

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Expression of Recom binant pEGFP·HPV16 E7 in Eukaryocyte

1. Virus Research Institution ofMedical College，Wuhan University Wuhan 430071，China

Abstract: Abstract：In order to make bases for construction of animal tumor model which expresses EGFP— HPV l 6E7，recombinant expression plasmid was constructed by techniques of gene recombination after screening an d identifing by restriction and PCR．Then the recombinant was tran sfected into mouse liver cancer cell by techniques of gene transfection and detected expression by fluoroscopy．pEGFP—HPV 1 6 E7 was identified by enzyme digestion an d PCR．Th e resent showed that the length ．inserted location and direction of the target gene was correct an d the expression of EGFP in tran sfected cell was observed Because of it’S pEGFP—HPV 1 6E7 expression plasmid that mak es it easy to assess the expression of EGFP-HPV 1 6E7 fusion protein and to sift the transfected cells．