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H5亚型禽流感病毒核蛋白基因重组质粒的构建
及其在Hela细胞的瞬时表达
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刘光亮1,2，王群1,2，童铁钢1,2，肖一红1,2，孟庆文1,2，吴东来1,2
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（1.中国农业科学院 哈尔滨兽医研究所，黑龙江 哈尔滨150001；
2.中国农业科学院 研究生院，北京100081）
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摘要：
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采用PCR技术从重组质粒pMel-NP扩增得到了禽流感病毒(AIV) A/Goose/Guangdong/1/96(H5N1)株的核蛋白(NP)基因，将其亚克隆至真核表达载体pVAX1上。将提取的pVAX-NP阳性克隆转染Hela细胞，48h后经间接免疫荧光试验和Western-blotting检测，NP基因在Hela细胞中已成功瞬时表达。表达产物的分子质量约为57ku，它与H5N1亚型AIV多克隆抗血清有较好的免疫反应。 |
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关键词：
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H5N1亚型禽流感病毒；NP基因；瞬时表达 |
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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)03-0189-04 |
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Construction and transient expression of recombinant
avian influenza virus with nucleoprotein gene |
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LIU Guang-liang1,2, WANG
Qun1,2,TONG Tie-gang1,2, XIAO
Yi-hong1,2,
MENG Qing-wen1,2, WU Dong-lai1,2 |
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(1. Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China;
2. School of Graduate, Chinese Academy of Agricultural Sciences, Beijing 100081, China) |
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Abstract: The NP gene of the AIV A/Goose/Guangdong/1/96(H5N1) strain was amplified from a recombinant plasmid
pMel-NP by PCR and subcloned into an eukaryotic expression vector pVAX1 and the recombinant vector was designated
pVAX-NP. The high quality pVAX-NP was prepared and the Hela cells were transfected. The recombinant vector
pVAX-NP was successfully expressed in the cells. Indirect immunofluorescence assay and
Western-blotting analysis showed that the nucleoprotein was expressed transiently in the Hela cells, which was 57ku in molecular weight, and had good immunologic reactivity with the polyclonal antiserum of AIV H5N1 subtype. |
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Key words: H5N1 subtype avian influenza virus；NP gene；transient expression |
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