HSV1-tk基因重组逆转录病毒载体的构建与表达

首都医科大学学报 ›› 1999, Vol. 20 ›› Issue (2): 83-85.

HSV1-tk基因重组逆转录病毒载体的构建与表达

李爱华¹, 刘天承², 田竟生²

1. 首都医科大学生化教研室;2. 首都医科大学神经科学研究所

收稿日期:1998-03-30 修回日期:1900-01-01 出版日期:1999-04-15 发布日期:1999-04-15

The Construction of pN₂A-HSV1-tk and Expression

Li Aihua¹, Liu Tiancheng², Tian Jingsheng²

1. Department of Biochemistry, Capital University of Medical Sciences;2. Beijing Institute for Neurosciences, Capital University of Medical Sciences

Received:1998-03-30 Revised:1900-01-01 Online:1999-04-15 Published:1999-04-15

摘要/Abstract

摘要： 通过DNA重组技术,将HSV1-tk基因片段重组到含有HSV1-tk启动子的pN₂A型逆转录病毒载体中。选取正向克隆的逆转录病毒重组DNA,以磷酸钙法转染ψ₂,PA317包装细胞后,经G418筛选,抗性克隆细胞上清能成功地感染NIH3T3,细胞系测定平均滴度为6.2×10⁵CFU/mL,最高可达1.5×10⁶CFU/mL.

关键词: HSV1-tk基因, 逆转录病毒载体, 基因转移

Abstract: The fulllength HSV1-tk gene was inserted into retrovirus vector pN₂Aby DNA recombinant techniques. The plasmid pN₂A-HSV1-tk was transfected into ψ₂, PA317 packaging cell line by DNA-calcium phosphate coprecipitation. The G418 resistant clones were selected and their medium could infect NIH3T₃ successfully and their average titer was 6.2×10⁵CFU/mL. The highest one reached 1.5×10⁶CFU/mL. The producer cell line will be used in gene therapy.

Key words: HSV1-tk gene, retrovirus vector, gene transfer

中图分类号:

Q784

李爱华;刘天承;田竟生. HSV1-tk基因重组逆转录病毒载体的构建与表达[J]. 首都医科大学学报, 1999, 20(2): 83-85.

Li Aihua;Liu Tiancheng;Tian Jingsheng. The Construction of pN₂A-HSV1-tk and Expression[J]. Journal of Capital Medical University, 1999, 20(2): 83-85.