猪2型圆环病毒Cap蛋白在伪狂犬病病毒中的表达  

Generation of the recombination PRV Tk^-/gE^-/ORF2^+ expressing PCV2 Cap protein

作　　者：宋云峰[1] 肖少波[1] 曹胜波[1] 张松林[1] 陈焕春[1] 金梅林[1] 

机构地区：[1]华中农业大学兽医学院动物病毒室,湖北武汉430070

出　　处：《中国兽医学报》2007年第2期155-158,共4页Chinese Journal of Veterinary Science

基　　金：国家"863"计划资助项目(2001AA213051)

摘　　要：用EcoR酶切伪狂犬病病毒疫苗株Tk-/gE-/LacZ+基因组,与含有猪2型圆环病毒(PCV2)ORF2基因的重组转移载体pIEORF2共转染IBRS-2细胞。收集病变细胞,经空斑纯化和PCR鉴定,获得了新的重组病毒Tk-/gE-/ORF2+。用Southern blot杂交证明该重组病毒构建正确,用间接免疫荧光法和Western blot证明该重组病毒可表达PCV2的主要免疫原性蛋白Cap蛋白。重组病毒在IBRS-2细胞上连续传15代后,遗传稳定。该重组病毒在IBRS-2细胞上增殖滴度为10-7.1TCID50/0.1mL,在鸡胚成纤维细胞上增殖滴度为10-4.8TCID50/0.1mL,在Marc-145细胞上增殖滴度为10-6.8TCID50/0.1mL。The genome of PRV vaccinal strain Tk-/gE+/LacZ+ was digested by EcoRⅠ,then co-transfected with transfer plasmid pIEORF2 into IBRS-2 cell line.The recombination virus was acquired by three times plaque purifications.PCR and Southern blot results showed that ORF2 gene has inserted into the expected location.Western blot and IFA results showed that the ORF2 gene can express in the recombinant PRV.TCID_50 of this recombination virus on different cell lines was measured to be IBRS-2 10-7.1,CEF 10-4.8,Marc-145 10-6.8 TCID_50/0.1 mL,respectively.

关 键 词：猪2型圆环病毒(PCV2) CAP蛋白 ORF2基因 疫苗 

分 类 号：S852.65[农业科学—基础兽医学]