犬瘟热病毒水貂分离株F1基因的克隆表达  

Cloning and Expression of F1 Gene of Mink CDV Isolate Strain

作　　者：张蕾[1] 柴秀丽[1] 张大鹏 张海玲[1] 赵建军[1] 高晗[1] 白雪[1] 徐磊[3] 闫喜军[1] 

机构地区：[1]中国农业科学院特产研究所,吉林吉林132109 [2]长春市动物卫生监督所,吉林长春130061 [3]中国兽医药品监察所,北京100081

出　　处：《中国畜牧兽医》2011年第9期65-68,共4页China Animal Husbandry & Veterinary Medicine

基　　金：科技部科研院所社会公益研究专项"野生动物重要传染病生态学发生;监测和控制技术"(2005DIB4J048)

摘　　要：本研究旨在利用基因工程的方法,制备犬瘟热(canine distemper,CD)疾病诊断用抗原。提取犬瘟热病毒基因组,RT-PCR扩增犬瘟热病毒(canine distemper virus,CDV)F1基因序列,克隆并测序。将F1基因定向克隆到原核表达载体pET28a多克隆位点,阳性重组质粒转化到大肠杆菌BL21(DE3)中,IPTG诱导表达。SDS-PAGE和Western blotting鉴定表达产物,并用Ni柱亲和层析纯化目的蛋白。结果显示表达的F1重组蛋白能与犬瘟热标准阳性血清发生反应,具有良好的反应原性,可以作为犬瘟热疾病的诊断用抗原。To prepare CDV antigen for diagnosis,gene engeneering method was explored.CDV genome was extracted,F1 gene was amplified by RT-PCR cloned and then sequenced.The F1 gene was cloned into expression plasmid pET28a and expressed by the induction of IPTG inducer.The recombined protein was examined by SDS-PAGE and Western blotting,purified by Ni column.The results indicated the recombined protein could react with CDV positive serum and might be used as CDV diagnostic antigen.

关 键 词：犬瘟热病毒 F1基因 表达 

分 类 号：Q78[生物学—分子生物学]