摘要
利用ＲＴ－ＰＣＲ克隆出共刺激分子人Ｂ７的全部编码ｃＤＮＡ，将其插入Ｅ１区替代的腺病毒载体ｐＡｘｌｃｗ，选择左向转录的ＣＤ８０重组腺病毒载体ｐＡｘｌｃｗ，ＣＩｈＣＤ８０，与经ＥｃｏＴ２２Ｉ酶切的Ａｄ５腺病毒ＤＮＧＡ－末端太复合物共转染２９３细胞，通过同源重组获得人ＣＤ８０的笔制缺陷性重组腺病毒，扩增到的病毒嘀度为４×１０＾９ｐｆｕ／ｍｌ．
The human CD80 full-length encoding cDNA was cloned by RT-PCR, and inserted into El-substituted aden-ovirus vector pAxlcw. Subsequently, the hCD80 recombinant adenovirus vector was cotransfected into 293 cells together with EcoT22I-digested Ad5-TPC, and the replication-deficient hCD80 recombinant adenovirus was generated efficiently by homologous recombination, with the tilers of 4×10~9pfu/ml. Infected with prepared hCD80 recombinant adenovirus in vitro, Hela cells expressed high levels of hCD80 48 hours after infection. These suggest that COS/TPC is an efficient method to prepare recombinant adenoviruses and the prepared hCD80 adenovirus could be used in cancer gene therapy.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
1997年第1期10-13,共4页
Chinese Journal of Cancer Biotherapy
基金
国家九五科技攻关项目(969060120)资助
关键词
共刺激分子
腺病毒载体
基因转移
肿瘤
costimulatory molecules
B7(CD80)
adenovirus vector
gene transfer
gene expression
