The Promoting Function of Long Terminal Repeat from  Reticuendotheliosis Virus

ZHAO Wen-ming; DING Jia-bo; JIANG Shi-jin; CUI Zhi-Zhong

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June 2004

ZHAO Wen-ming, DING Jia-bo, JIANG Shi-jin and CUI Zhi-Zhong. The Promoting Function of Long Terminal Repeat from Reticuendotheliosis Virus[J]. Virologica Sinica, 2004, 19(3): 255-258.

Citation: ZHAO Wen-ming, DING Jia-bo, JIANG Shi-jin, CUI Zhi-Zhong. The Promoting Function of Long Terminal Repeat from Reticuendotheliosis Virus .VIROLOGICA SINICA, 2004, 19(3) : 255-258.

禽网状内皮组织增殖病病毒LTR序列的启动子功能*

1.
1. 扬州大学畜牧兽医学院, 江苏扬州 225009
2.
山东农业大学动物科技学院, 山东泰安 271018

通讯作者： 赵文明, ;

摘要

通过PCR方法，将禽网状内皮组织增殖病病毒（REV）的长末端重复序列（LTR）扩增并克隆进pUC-18质粒多克隆位点（MCS）的EcoR I和Sac I之间，并以BGH基因的多聚腺苷酸序列作为终止子克隆到Sph I和Hind Ⅲ之间，构建成重组质粒pUC-LTR。将GFP基因和REV囊膜糖蛋白gp90基因分别克隆到pUC-LTR载体中，获得质粒pUC-LTR-GFP和质粒pUC-LTR-gp90。重组质粒经转染48h，能够检测到外源基因的表达。本研究提示，REV LTR能够作为启动子构建表达质粒。
- 禽网状内皮组织增殖病病毒（REV）
- , 长末端重复序列（LTR）
- , 启动子
- , 表达质粒

The Promoting Function of Long Terminal Repeat from Reticuendotheliosis Virus

1.
1. College of Animal Husbandy and Veterinary Medicine, Yangzhou University, Yangzhou 22500, China
2.
College of Animal Science and Technology, Shandong Agricultural University, Taian 271018, China

Corresponding author: ZHAO Wen-ming,

Abstract

The long terminal repeat (LTR) of Reticuendotheliosis virus (REV) was amplified by PCR technique and cloned into pUC18 vector at the sites of EcoRI and SacI, and the BGH polyadenylation signal sequence was cloned at the sites of Sph I and Hind Ⅲ as a terminor. The positive clone was named pUC-LTR, which was used as a basic expressing vector to validate the activity of LTR. Green fluorescent protein (GFP) and REV envelope glycoprotein 90 (gp90) gene was cloned at the downstream of LTR in the pUC-LTR vector respectively as a reporter. These two recombinants pUC-LTR-GFP and pUC-LTR-gp90 were then transfected into Chicken Embryo Fibroblast (CEF) cells. 48h after the transfection, we could detect the expression of GFP and gp90. This study shows the LTR sequence could be used as a promoter to construct expressing vectors.
- Reticuendotheliosis virus (REV)
- , Long terminal repeats (LTR)
- , Promoter
- , Expressing vector

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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

The Promoting Function of Long Terminal Repeat from Reticuendotheliosis Virus

Corresponding author: ZHAO Wen-ming,

1. 1. College of Animal Husbandy and Veterinary Medicine, Yangzhou University, Yangzhou 22500, China
2. College of Animal Science and Technology, Shandong Agricultural University, Taian 271018, China

Abstract: The long terminal repeat (LTR) of Reticuendotheliosis virus (REV) was amplified by PCR technique and cloned into pUC18 vector at the sites of EcoRI and SacI, and the BGH polyadenylation signal sequence was cloned at the sites of Sph I and Hind Ⅲ as a terminor. The positive clone was named pUC-LTR, which was used as a basic expressing vector to validate the activity of LTR. Green fluorescent protein (GFP) and REV envelope glycoprotein 90 (gp90) gene was cloned at the downstream of LTR in the pUC-LTR vector respectively as a reporter. These two recombinants pUC-LTR-GFP and pUC-LTR-gp90 were then transfected into Chicken Embryo Fibroblast (CEF) cells. 48h after the transfection, we could detect the expression of GFP and gp90. This study shows the LTR sequence could be used as a promoter to construct expressing vectors.