摘要
为建立1株猪源2009/H1N1流感病毒A/swine/Heilongjiang/44/2009(HLJ44)的反向遗传系统,利用双向表达质粒p HW2000,分别构建了该病毒株8个基因节段的重组质粒,将其共转染于293T和MDCK混合培养的细胞,拯救出重组病毒R-HLJ44。测序结果表明,R-HLJ44与亲本病毒HLJ44的核苷酸序列完全一致;二者在细胞上具有相似的增殖特性;抗原性未发生变化;分别以106TCID50的剂量鼻腔感染BALB/c小鼠,结果显示R-HLJ44与亲本HLJ44在小鼠脏器中的复制滴度也基本一致。以上结果表明拯救的重组病毒保持了与亲本病毒一致的生物学特性。该病毒反向遗传系统的建立,为进一步研究H1N1亚型流感病毒的致病分子机制及新型疫苗研制等奠定了基础。
To establish the reverse genetic system of a pandemic 2009 / H1N1 influenza virus,we firstly constructed the recombinant plasmids containing the eight gene segments of A / swine / Heilongjiang / 44 / 2009（ HLJ44） based on the p HW2000 plasimds,respectively. Then the recombinant virus（ designated R-HLJ44） was rescued by co-transfecting the eight recombinant plasmids into the cell mixtures of 293 T and MDCK. Whole genome sequence analysis indicated that the R-HLJ44 had the similar antigenic characteristics with the parental virus of HLJ44 and no nucleotides were changed compared with HLJ44. In addition,the virus replication titer of R-HLJ44 in BALB / c mice was similar to the parent virus by infecting mice with 106TCID50 of viruses. These results indicated the reverse genetic system of 2009 / H1N1 influenza virus was established,which lays the foundation for the further study on the pathogenicity mechanism and novel vaccine development of H1N1 subtype influenza viruses.
作者
许榜丰
孟沙沙
吴运谱
陈艳
杨焕良
乔传玲
陈化兰
XU Bangfeng MENG Shaha WU Yunpu CHEN Yan YANG Huanliang QIAO Chuanling＊ CHEN HuMan(Animal Influenza Laboratory of the Ministry of Agricultural, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China)
出处
《畜牧与兽医》
北大核心
2017年第1期61-64,共4页
Animal Husbandry & Veterinary Medicine
基金
哈尔滨市科技创新人才项目(2014RFXYJ116)
