基于甲病毒复制子的犬瘟热核酸疫苗的构建及表达性研究  

Study on the Construction and Expression Characteristics of Canine Distemper DNA Vaccine Based on SFV-replicon

作　　者：孙建涛[1,2] 侯绍华[2] 于萍[2,3] 朱鸿飞[2] 梁欠 郝志明[1] 

机构地区：[1]天津农学院动物科学系,天津300384 [2]中国农业科学院北京畜牧兽医研究所,北京100193 [3]中国农业科学院兰州畜牧与兽药研究所,甘肃兰州730050

出　　处：《中国畜牧兽医》2013年第9期96-100,共5页China Animal Husbandry & Veterinary Medicine

基　　金：农业部动物疫情监测与防治项目;863动物疫病分子诊断技术研究与产品创制项目(2012AA101302);国家科技支撑计划重点项目(2010BAD04B02);公益性行业(农业)科研专项经费项目(200903027)

摘　　要：本试验利用T-A克隆技术,构建克隆载体pEASY-Blunt-F,BamHⅠ酶切pEASY-Blunt-F,回收F基因,将其克隆至pSCA1真核表达载体中,获得重组质粒pSCA1-F。经DNA测序、限制性内切酶分析和PCR鉴定,结果表明重组质粒pSCA1-F成功构建。通过间接免疫荧光试验和Western blotting证实F基因在转染细胞中表达。此外,细胞凋亡的检测结果表明,pSCA1-F能引起转染的细胞发生凋亡。The clone vector pEASY-Blunt-F with fusion gene of canine distemper virus（CDV） isolated from vaccine strain was successfully constructed by T-A clone technique and was digested with restriction enzyme of BamHⅠ. The F gene was cloned into eukaryotic expression vector pSCA1. Finally a recombinant plasmid named pSCA1-F was constructed and identified by PCR, restriction enzyme analysis and sequencing. pSCA1-F was transfected into BHK-21 cells by using X-tremeGENE 9 DNA Transfection Reagent,IFA and Western blotting were performed to detect F expressions,the results showed that F gene had expressed in the transfected cells.Furthermore,apoptosis assay confirmed that pSCA1-F could induce the transfected cells apoptosis.

关 键 词：核酸疫苗 犬瘟热病毒 F基因 甲病毒复制子 细胞凋亡 

分 类 号：Q786[生物学—分子生物学]