H5N1亚型高致病性禽流感病毒A/duck/HuBei/49/05株反向基因操作系统的建立  

Establishment of Highly Pathogenic H5N1 Avian Influenza Virus A/duck/HuBei/49/05 Base on Reverse Genetics System

作　　者：苏胜杰[1] 许静[2] 宋家升[2] 李雁冰[2] 希尼尼根[1] 

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010018 [2]中国农业科学院哈尔滨兽医研究所农业部动物流感重点开放实验室兽医生物技术国家重点实验室,黑龙江哈尔滨150001

出　　处：《畜牧与饲料科学》2010年第2期20-21,27,共3页Animal Husbandry and Feed Science

摘　　要：水禽是禽流感病毒的天然储存库,为了研究高致病性禽流感病毒对水禽致病性的分子基础,研究构建了DKHB/49/05的8质粒反向遗传操作系统。用Trizol从含A/duck/HuBei/49/05株病毒的鸡胚尿囊液中提取总RNA,用RT-PCR扩增PB2、PB1、PA、HA、NP、NA、M、NS基因片段,将其分别克隆至pBD载体中。利用8质粒反向遗传操作系统,将重组的pBD质粒共转染293T细胞,成功拯救了该病毒,并命名为R-DKHB。Waterfowl are the natural repository of avian influenza virus.To study the pathogenic molecular basis of highly pathogenic avian influenza from waterfow,8 recombinant PBD plasmids and rescued the recombinant virus were constructrd by reverse genetics techniques.The total RNA was extracted from the allantoic fluids of chick embryo with the infection of A/duck/HuBei/49/05 virus,PB2,PB1,PA,HA,NP,NA,M and NS were amplified by RT-PCR respectively,and they were cloned into PBD vector.Moreover,they also were transfected into 293 T cells.The result indicated that the recombinant virus was successfully rescued and named as R-DKHB.

关 键 词：H5N1 禽流感病毒 反向遗传 

分 类 号：S852.659.5[农业科学—基础兽医学] Q785[农业科学—兽医学]