摘要
为研究重组腺病毒接种实验动物后的免疫反应性，利用ＲＴ－Ｐ Ｃ Ｒ方法，从ＨＡＶ的ＲＮＡ中克隆了结构蛋白基因插入穿梭质粒ｐＸＣＸ２Ｎｏｔ Ｉ，通过磷酸钙－ＤＮＡ共 沉淀技术，将复制缺陷型腺病毒载体与线形化的ｐＸＣＸ２－ＣＭＶ－ＨＡＶ共转染２９３细胞。一系列检测方法证明产生了重组腺病毒ｒＡｄＨＡＶ。纯化后的ｒＡｄＨＡＶ滴度为１×１０９ＴＣＩＤ５０／ｍＬ ，腹腔注射免疫昆明种小白鼠后，可诱导产生抗ＨＡＶ ＩｇＧ和ＨＡＶ中和抗体。复制缺陷型腺病毒 可作为发展基因工程病毒疫苗载体的有效系统。
In order to investigate the immunity of the recombinant a denoviruses inoculated to the animals, structural gene from RNA of HAV was clone d by RT-PCR and inserted into the shuttle plasmid(pXCX2Not I). A replication－defective adenovirus was rescued in 293 packing cells via homologous recombina tion of both plasmid pXCX2-CMV-HAV and pJM17 containing Ad5 genome with dele tions of E1 region by calcium phosphate technique. A series of methods were employed to identify the generated recombinant ademovirus (rAdHAV). The titer of rAd HAV stocks was up to 1×109TCID50/mL. The Kunming mice were administrate d with rAdHAV by intraperitonea injection(i.p.). Anti-HAV IgG and HAV neutraliz ing antibodies were induced. In conclusion, a recombinant replication-defective adenovirus is an efficient delivery system to develop recombinant virus vaccine .
出处
《中国病毒学》
CSCD
2002年第1期51-55,共5页
Virologica Sinica
关键词
重组腺病毒
腹腔注射
免疫反应
Recombinant adenovirus
Intraperitonea infection(i.p.)
Im une response
