从cDNA感染性克隆产生冷适应重配A型人流感病毒和免疫原性的初步研究.pdf

24 4 20087 IMMUNOLOGICAL JOURNAL Vol. 24 No. 4 Jul. 2008 # 373 # [ ] 1000-8861(2008)04-0373-05 cDNA A 1,2 2 2 1 1 3 2 1* 叶 艺 ,杨鹏辉 , 罗德炎 , 吴 朔 , 周 朋 , 刘秀梵 , 王希良 ,于三科 ( 1. 西北农林科技大学 动物医学 ,陕西杨陵712100;2.军事医学科学 微生物流行病研究所病原微生物生物安全国家重点实验 室, 北京 100071;3. 扬州大学农业部畜禽传染病学重点开放实验室, 江苏 扬州225009) [ ] cDNA A , RT-PCR, 2006 2007 APNew CaledoniaP20P99 HANA , pMDV-A-HApMDV-A-NA, 26 COS-1, A , A , , [] ;; ;; [] R183.3 [ ] A Generation of a cold-adapted reassortantinfluenza A virus from cDNA and research on its immuno- genicity YEYi, YANG Peng-hui, LOU De-yan, WU Shuo, ZHOU Peng, LIU Xiu-fan, WANG X-i liang, YU San-ke (College of Veterinary Medicine , N orthwest AF University , Yangling 712100, China) [Abstract] Objective To generate the cold-adapted reassortant influenza A virus by reverse genetics from cDNAs and to analyze its phenotype characters and immunogenicity. Methods The hemagglutinin (HA) and neuramindase ( NA) genes of APNew CaledoniaP20P99 strain were amplified by RT-PCR, and then cloned intothe plasmid pAD3000. The constructed bidirectional expression vectors, pMDV-A-HA and pMDV-A-NA, weretransfected into the COS-1 cell togetherwith other6 plasmids from cold-adapted virus recovery system. The phenotype characters and immunogenicity of the reassortant influenzaA virus wereanalyzed. Results Thereassortant influenzaA virus was recovered with cold-adapted phenotypeandtemperature sensitivity. And its bionomics and immunogenicitywas analyzed. Conclusion Thereassortant influen- za A virus with cold-adapted phenotype and temperaturesensitivity is recovered successfully and its immunogenicity is determined, which lays thefoundation for recovering the human influenza attenuated strain and developing the influenza attenuated vaccine. [ ey words] Reversegenetics