| H7亚型禽流感病毒血凝素基因的真核表达载体的构建及鉴定 |
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| 引用本文： | 杨旭芹,梁光军,张小荣,文其乙,刘秀梵. H7亚型禽流感病毒血凝素基因的真核表达载体的构建及鉴定[J]. 中国家禽, 2004, 8(Z1): 31-34 |
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| 作者姓名： | 杨旭芹 梁光军 张小荣 文其乙 刘秀梵 |
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| 作者单位： | 扬州大学兽医学院畜禽传染病学农业部重点开放实验室,江苏扬州225009 |
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| 摘 要： | 参考已发表的H7亚型禽流感病毒(AIV)血凝素(HA)基因序列设计引物,以pUCH7为模板,经PCR扩增出一条1.7kb的HA全基因片段,将该片段定向克隆到真核表达质粒载体pcDNA3.1(一)中,转化大肠埃希氏菌DH5α,小量制备重组质粒pcDNA-HA,酶切鉴定正确后,转染COS-1细胞,经免疫荧光鉴定其体外表达情况。结果表明H7亚型AIV HA在COS-1细胞中获得了成功表达。用该重组质粒pcDNA-HA免疫BALB／C小鼠,制备免疫血清,经免疫印迹实验证实该H7亚型AIV HA在小鼠体内也得到了良好的表达。
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| 关 键 词： | H7亚型禽流感病毒 血凝素基因 真核表达质粒载体 表达 |
| 修稿时间： | 2004-10-19 |
Construction and Identification of Hemagglutinin Gene of Avian Influenza Virus Subtype H7 Eukaryotic Expression Vetor |
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| Yang Xuqin,Liang Guangjun,Zhang Xiaorong,Wen Qiyi,Liu Xiufan. Construction and Identification of Hemagglutinin Gene of Avian Influenza Virus Subtype H7 Eukaryotic Expression Vetor[J]. China Poultry, 2004, 8(Z1): 31-34 |
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| Authors: | Yang Xuqin Liang Guangjun Zhang Xiaorong Wen Qiyi Liu Xiufan |
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| Abstract: | One 1.7 kb DNA fragment was amplified by PCR from pUCH7.The HA gene of H7 subtype AIV A/Afri. Star./Eng-Q/983/79(H7N1)was inserted into eukaryotic expression vector pcDNA3.l (-).The recombinant plasmid pcD-NA-HA was transfected into COS-1 cell by lipinfectin reagent after it was identified by restriction digestion.The result of IFA revealed the HA gene was expressed successfully in COS-1 cell.The BALB/C mice had been immunized by recombinant plasmid pcDNA-HA.And the result of Western Blot revealed the HA gene was also expressed successfully in mice and generated antibody against HA of H7 subtype AIV. |
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| Keywords: | H7 subtype AIV hemagglutinin gene eukaryotic expression vetor expression |
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