摘要
血凝素(HA)和神经氨酸酶(NA)是禽流感病毒(AIV)的两种表面糖蛋白,主要诱导机体产生体液免疫应答。本研究采用SDS-蛋白酶K法提取禽流感病毒分离株A/Goose/Guangdong/3/96(H5N1)的SPF鸡胚尿囊毒RNA,RT-PCR扩增完整的NA cDNA基因,将其克隆到载体pSY538中的禽痘病毒早晚期启动子 LP2EP2的下游,然后再将含有启动子的 NA基因片段亚克隆到已有的AIVHA基因重组禽痘病毒转移载体pSY(HA+LacZ)中,构建了HA、NA双重组禽痘病毒转移载体pSY(HA+NA+LacZ),将它与禽痘病毒疫苗株S-FPV-017共转染鸡胚成纤维细胞,在X-gal存在的条件下,经蓝斑筛选、六选蚀斑纯化、PCR鉴定等,结果表明已获得一株能同时稳定表达HA和NA蛋白的重组禽痘病毒,为禽流感基因工程病毒活载体疫苗的研究奠定了基础。
Haemagglutinin and neuraminidase are two kinds of surface glycoprotein, which primarily induce humoral immunity. In this study, avian influenza virus isolate A/Goose/Guangdong /3/96((H5N1) was propagated in SPF chinken embryo. Virus RNA was extracted from allantoic fluid by means of SDS-protease K. The full-length NA cDNA was amplified using RT-PCR,then cloned into vector pSY538, which con- taining fowlpox virus early/late promoter LP2EP2.The fragment of NA promoted by LP2EP2 was subcloned into pS...
出处
《中国预防兽医学报》
CAS
CSCD
2000年第S1期178-,共1页
Chinese Journal of Preventive Veterinary Medicine
基金
973项目资助!(G1999011902
G1999011905)
关键词
禽流感病毒
血凝素基因
神经氨酸酶基因
重组禽痘病毒
Avian Influenza Virus
Haemagglutinin gene
Neuraminidase gene
Recombinant Fowlpox Virus