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

doi:10.12160/j.issn.1672-5190.2010.02.008

畜牧与饲料科学 ›› 2010, Vol. 31 ›› Issue (2): 20-20.doi: 10.12160/j.issn.1672-5190.2010.02.008

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

苏胜杰[1] 许静[2] 宋家升[2] 李雁冰[2] 希尼尼根[1]

[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

出版日期:2010-02-20 发布日期:2010-02-20
通讯作者: 苏胜杰
作者简介:苏胜杰（1984-），男，硕士，主要研究方向为禽流感分子病毒学。通讯作者：希尼尼根（1970-），男，副教授，博士，主要研究方向为动物传染病学。

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

SU Sheng-jie,XU Jing ,SONG Jia-sheng,LI Yan-bing,Xininigen（1.College of Veterinary Science,Inner Mongolia Agricultural University,Hohhot 010018,China;2.Animal Influenza Laboratory of the Ministry of Agriculture and National Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute CAAS,Harbin 150001,China）

Online:2010-02-20 Published:2010-02-20

摘要/Abstract

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。

Abstract: 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.

中图分类号:

S852.659.5 Q785

苏胜杰,许静,宋家升,李雁冰,希尼尼根. 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[J]. 畜牧与饲料科学, 2010, 31(2): 20-20.

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

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