华南农业大学学报2011，Vol.32Issue(2)：80-84,92,6.
双表达H9N2禽流感病毒HA基因的假型重组杆状病毒系统的构建
Construction of A Pseudotype Baculovirus Dual Expression System-Based Avian Influenza H9N2 Virus HA Gene
摘要
Abstract
HA gene of avian influenza H9N2 virus amplified by PCR from the recombinant plasmid pMD18-HA was sub-cloned into pBACsurf-1. The fusion gene containing HA and gp64 was inserted into pcDNA3.1 ( + ) to construct the recombinant plasmid pcDNA-H9-HA. A fragment of the CMV promoterH9HA-gp64 fusion gene cassette was excised from pcDNA-H9-HA and inserted into the baculovirus transfer vector pFastBac-G to construct the recombinant plasmid pFastBac-G-H9-HA. The plasmid was transformed into Escherichia coli DH10Bac competent cells to obtain recombinant shuttle vector Bacmid-G-H9-HA. The recombinant baculovirus BV-DuaI-H9-HA was generated in Sf9 cells by cotransfection of the recombinant shuttle vector Bacmid-G-H9-HA with LipofectamineTM2000. Indirect immunofluoresent assay demonstrated that BV-DuaI-H9-HA efficiently transducted the mammalian cells in vitro and HA gene was successfully expressed. Immunoelectron microscopy showed that HA protein was surface-displayed on the BV-DuaI-H9-HA virus.关键词
禽流感病毒/H9N2亚型/双表达/HA基因/重组假型杆状病毒Key words
avian influenza virus/ H9N2 subtype/ dual expression/ HA gene/ pseudotype baculovirus分类
农业科技引用本文复制引用
林文耀,樊惠英,程晓亮,陈筱薇,叶煜,廖明..双表达H9N2禽流感病毒HA基因的假型重组杆状病毒系统的构建[J].华南农业大学学报,2011,32(2):80-84,92,6.基金项目
国家自然科学青年基金(30800826) （30800826）
广东省博士启动基金(845106420l001131) （845106420l001131）
农业微生物学重点实验室开发课题(20090010) （20090010）
华南农业大学校长基金(5500-K08240) （5500-K08240）
