| H9N2亚型禽流感病毒血凝素基因的克隆及原核表达 |
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| 引用本文： | 李春红,董玉龙.H9N2亚型禽流感病毒血凝素基因的克隆及原核表达[J].安徽农业科学,2009,37(18):8375-8376. |
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| 作者姓名： | 李春红 董玉龙 |
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| 作者单位： | 河北北方学院动物医学系,河北张家口,075131;河北北方学院理学院,河北张家口,075131 |
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| 摘 要： | 据GenBank收录的H9N2亚型禽流感病毒血凝素（HA）基因序列设计并合成引物，以H9N2亚型禽流感病毒RNA为模板，用R1PCR方法扩增了预计约1700bp的HA基因，将此扩增产物克隆进pMD18-T载体，采用限制性酶切及序列测定鉴定阳性重组克隆子结果表明HA基因长为1683bp。基于HA信号肽在表达中的负作用，研究通过基因工程手段缺失HA蛋白位于起始的信号肽的编码户列，获得了缺失HA蛋白信号肽的HA基因，将其亚克隆到pGEX-KG中，与GST融合表达。SDS-PAGE显示：融合表达的蛋白分子量乡为90kDa。
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| 关 键 词： | 禽流感病毒 血凝素基因 原核表达 |
Clone and Procaryotic Expression of Hemagglutinin Gene of H9N2 Avian Influenza Virus |
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| Institution: | LI Chun-hong et al (Department of Animal Medicine, Hebei Northern University, Zhangjiakou,Hebei 075131 ) |
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| Abstract: | According to HA gene sequence of H9N2 avian influenza virus collected by GenBank,We designed and synthesized primers,1 700 bp fragment was amplified from H9N2 AIV RNA by RT-PCR and cloned into the pMD18-T vector.Restrictive enzyme and sequence were used to determinate positive recombinant cloning.The results showed that HA gene consisted of 1 683 bp.Based on the negative function of signal peptide of HA gene in expression,we lacked the signal peptide of HA gene which lies in initial coding sequence through the genetic engineering means and obtained the HA gene which lacked signal peptide,then partial fragment of HA was subcloned into prokaryotic expression vector pGEX-KG,and was fusion expressed with GST.SDS-PAGE showed that protein molecular weight was about 90 kDa. |
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| Keywords: | Avian Influenza virus Hemagglutinin(HA)gene Prokaryotic expression |
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