首都医科大学学报 ›› 2010, Vol. 31 ›› Issue (6): 695-700.

• HN/ADS基础与临床研究进展 • 上一篇    下一篇

gp120转基因小鼠制备及初步鉴定

张洪海, 孙玉, 柳雅立, 张彤, 吴昊, 陈德喜   

  1. 首都医科大学附属北京佑安医院性病艾滋病实验室
  • 收稿日期:2010-08-09 修回日期:1900-01-01 出版日期:2010-12-24 发布日期:2010-12-24
  • 通讯作者: 陈德喜

Preparation and Identification of gp120 Transgenic Mice

ZHANG Hong-hai, SUN Yu, LIU Ya-li, ZHANG Tong, WU Hao, CHEN De-xi   

  1. STD & AIDS Laboratory, Beijing Youan Hospital, Capital Medical University
  • Received:2010-08-09 Revised:1900-01-01 Online:2010-12-24 Published:2010-12-24

摘要: 目的 构建HIV-1B'亚型gp120转基因载体,制备gp120转基因小鼠。方法 扩增gp120全长序列,根据读码框架定向连接到pSEAP2-control外分泌性真核表达载体上,插入GFAP启动子,回收含启动子-gp120-增强子的功能片段制备转基因小鼠,PCR和RT-PCR验证基因整合与mRNA转录。结果 酶切鉴定与测序验证均证明构建了受GFAP启动子调控的gp120转基因载体,蛋白印迹分析显示其分泌性和组织特异性都很高;RT-PCR证实转基因小鼠存在gp120mRNA转录。结论 成功构建了gp120转基因载体,并生产了founder小鼠。

关键词: 人类免疫缺陷病毒, gp120, 痴呆, 动物模型

Abstract: Objective To construct HIV-1 B' subtype gp120 transgenic vector and prepare gp120 transgenic mice.Methods gp120 gene was amplified and cloned to pSEAP2-control vector according to reading frame,then GFAP promoter was inserted.The function fragment with promoter-gp120 gene-enhancer was digested and extracted to prepare transgenic mice.The gene integration and mRNA transcription was validated by PCR and RT-PCR.Results GFAP promoter controlled gp120 transgenic vector was successfully constructed and validated by gene sequencing,Western blotting analysis showed that the secretion and tissue specificity of gp120 was high enough;The existence of gp120 mRNA transcription in transgenic mice was confirmed by RT-PCR.Conclusion gp120 transgenic vector was successfully constructed and founder mice were produced.

Key words: human immunodeficiency virus, gp120, dementia, animal model

中图分类号: