复制缺陷型腺病毒AdE1CMV-NGF的制备

首都医科大学学报 ›› 2001, Vol. 22 ›› Issue (1): 24-26.

复制缺陷型腺病毒AdE1CMV-NGF的制备

刘世全¹, 张薇², 贾东辉³, 王申五¹, 郑姝颖¹, 王薇⁴, 张惠蓉⁴

1. 北京医科大学人民医院中心实验室;2. 北京友谊医院眼科;3. 中国医科院肿瘤医院;4. 北医大三院眼科

收稿日期:2000-01-29 修回日期:1900-01-01 出版日期:2001-01-15 发布日期:2001-01-15

The Preparation of Replication-deficient Recombinant Adenovirus AdE1CMV-NGF

Liu Shiquan¹, Zhang Wei², Jia Donghui³, Wang Shenwu¹, Zheng Shuying¹, Wang Wei⁴, Zhang Huirong⁴

1. Center Laboratory, People's Hospital, Beijing Medical University;2. Beijing Friendship Hospital;3. Chinese Academy of Medical Science;4. The Third Clinical College, Beijing Medical University

Received:2000-01-29 Revised:1900-01-01 Online:2001-01-15 Published:2001-01-15

摘要/Abstract

摘要： 通过DNA重组技术,经亚克隆和克隆过程构建分泌表达型重组腺病毒穿梭质粒pAdE1CMV-NGF;制备完整高纯度辅助病毒pJM17.将pAdE1CMV-NGF和pJM17在293细胞中同源重组,获得复制缺陷型腺病毒重组体AdE1CMV-NGF.梯度系列稀释测得滴度为4.6×10¹⁰PFU/mL.

关键词: NGF基因, 腺病毒载体, 基因转移, 同源重组

Abstract: With the DNA recombinant techniques, we constructed a secretable form of adenoviral shuttle plasmid pAdE1CMV-NGF through sub-cloning and cloning steps. And then pAdE1CMV-NGF was cotransfected with pJM17 in 293 cells with the application of FuGENE 6 to produce the replication deficient virions containing the cDNA of interest, AdE1CMV-NGF. The titer reached 4.6×10¹⁰ PFU/mL by grade-dilution evaluation.

Key words: NGF gene, adenovirus vector, gene transfer, homologous recombination

中图分类号:

刘世全;张薇;贾东辉;王申五;郑姝颖;王薇;张惠蓉. 复制缺陷型腺病毒AdE1CMV-NGF的制备[J]. 首都医科大学学报, 2001, 22(1): 24-26.

Liu Shiquan;Zhang Wei;Jia Donghui;Wang Shenwu;Zheng Shuying;Wang Wei;Zhang Huirong. The Preparation of Replication-deficient Recombinant Adenovirus AdE1CMV-NGF[J]. Journal of Capital Medical University, 2001, 22(1): 24-26.

[1]	方红娟;杨金奎. 大鼠ACE2腺病毒载体的构建及其在INS-1细胞中的表达[J]. 首都医科大学学报, 2009, 30(4): 449-454.
[2]	肖新如;康熙雄;赵继宗. BAI1的血管生成抑制作用及肿瘤生长抑制作用的研究[J]. 首都医科大学学报, 2006, 27(3): 351-354.
[3]	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.