摘要
从pMD18-T-HA阳性质粒中扩增出H7亚型禽流感病毒(AIV)HA1基因,并亚克隆至昆虫杆状病毒转移载体pBlueBacHis2A,将筛选的重组质粒命名为pBlueBacHisH7-HA1,与线性化杆状病毒DNA(Bac-N-BlueDNA)共转染sf9昆虫细胞,挑取蓝色蚀斑,经3次蚀斑纯化,得到重组杆状病毒rpBlueHisH7HA1。Western-blotting和间接免疫荧光试验结果显示,HA1在昆虫细胞中得到表达,且表达产物具有抗原性。抗原特异性试验证明,重组HA1蛋白与鸡H5、H9亚型AIV抗血清不发生交叉反应。血凝试验证明,重组HA1蛋白具有良好的血凝性。
The HA1 gene of avian influenza virus H7 subtype was amplified from recombinant plasmid pMD-18-T-HA by PCR. The PCR product was subcloned into pBlueBacHis2A vector. The recombinant transfer vector pBlueHisH7-HA1 was screened at random. After pBlueHisH7-HA1 was co-transfected with Bac-N-Blue DNA by cationic liposome-mediated transfection, and was purified by three cycles of plaque assay with chromogenic substracte X-gal in low temperature-melting agarose, a recombinant baculovirus, designated as rpBlueHisH7-HAl,was obtained. Western-blotting and indirect immunofluorescence tests showed the HA1 gene was well expressed,and the expressed product had antigenicity. An antigen specific test showed that the recombinant did not have any cross-reaction with chicken H5 and H9 antisera. A hemagglutinin test certified that it had good hemagglutination.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2007年第3期232-235,共4页
Chinese Veterinary Science
基金
国家科技攻关计划项目(2004BA519A23)
