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H9N2禽流感病毒HA2基因重组杆状病毒的表达
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王彬，唐志芬，齐岩，孙凌霜，王君伟
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（东北农业大学 动物医学院，黑龙江 哈尔滨150030）
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摘要：
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从pMD18-T-HA阳性质粒扩增了H9N2亚型禽流感病毒的HA2基因，将扩增到的HA2基因克隆至昆虫杆状病毒转移载体 pBlueBacHis2A中。将其与杆状病毒共转染于Sf9昆虫细胞，经蚀斑筛选纯化重组杆状病毒，用其感染Sf9昆虫细胞，并优化表达条件。SDS-PAGE和Western-blotting分析表明，表达产物的分子质量约为27ku。Dot-ELISA分析表明，表达的HA2融合蛋白可与鸡抗H9N2亚型血清发生特异性反应，而与H5和H7亚型抗血清间无交叉反应。 |
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关键词：
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禽流感病毒；H9N2亚型；HA2基因；重组杆状病毒；抗原性 |
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中图分类号：S 852.659.5：Q 786 文献标识码：
A 文章编号：1673-4696(2006)05-0362-04 |
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Recombinant baculovirus expression of HA2 gene of
avian influenza virus H9N2 subtype |
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WANG Bin，TANG Zhi-fen，QI Yan，SUN Ling-shuang，WANG Jun-wei |
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(College of Veterinary Medicine, Northeast Agricultural University,Harbin 150030, China) |
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Abstract: Using a pair of specific primers designed according to the pMD18-T-HA, the HA2 gene of avian influenza virus H9N2 subtype was amplified by PCR. The PCR product was cloned into pBlueBacHis2A to obtain an eukaryotic recombinant plasmid pBlueBacHis2A-HA2, and then Sf9 cells were transfected with the recombinant plasmid. After plaque screening, the target HA2 gene was expressed in insect cell Sf9, and the expressed protein was 27ku in molecular weight. Western-blotting and Dot-ELISA analysis showed that the recombinant protein had good reactivity against H9N2 subtype serum and did not have any cross-reaction with chicken H5 and H7 antiserum. |
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Key words: avian influenza virus; H9N2 subtype; HA2 gene; recombinant baculovirus; antigenicity |
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