摘要
利用反向遗传技术将含有A／Chicken／Shanghai／F／98（H9N2）株禽流感病毒（avian influenza virus，AIV）的6个内部基因与H5N1亚型AIV的2个表面基因HA和NA共转染COS-1细胞，产生了6＋2全禽源的重配AIV。将H5N1亚型AIV的HA基因经基因突变致弱，然后将A／Chicken／Shanghai／F／98（H9N2）AIV的6个内部基因的cDNA和以上致弱的禽源HA基因及NA基因的cDNA分别克隆到转录／表达载体pHW2000中，构建成8个转录／表达质粒。将8个质粒共转染COS-1细胞，24h后收获细胞及上清接种SPF鸡胚，72～90h后鸡胚死亡，收取鸡胚尿囊液进行血凝、血凝抑制试验、序列分析、病毒致病性试验和动物免疫保护试验，最终证实产生了致弱的全禽源AIV瘩苗候诜株．
Eight-plasmid system was used for the generation of avian influenza virus （AIV） vaccine candidate strain rgF-H5△N1 AIV. The strain contained hemagglutinin （HA） and neuraminidase（ NA ） genes from epidemic strains in a background of internal genes derived from A/Chicken/Shanghai/F/98 （H9N2） strain. The removal of the multibasic amino acid motif in the HA gene associated with high pathogenicity in chickens. The rgF-H5△N1 AIV attenuated the viral virulence for chickens without altering the antigencity of the HA. A for- malin-inactivated vaccine prepared from this virus was immunogenic and protected chickens from subsequent wild-type H5N1 virus challenge. This is the first successful attempt to develop an H5N1 vaccine from avian influenza genes by plasmid-based reverse genetics at home and abroad.
出处
《病毒学报》
CAS
CSCD
北大核心
2006年第2期126-130,共5页
Chinese Journal of Virology
基金
江苏省属高校重大基础研究项目资助(05KJA23016)
农业部资助项目(农办牧200183)
关键词
反向遗传技术
AIV
质粒
转染
reverse genetics method
avian influenza virus（AIV）
plasmid
vaccine
transfection
