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当前位置: 首页 > 文章 > H9N2亚型禽流感病毒HA基因的表达及其单因子血清的制备 中国畜牧兽医 2015,42 (11) 2888-2894
Position: Home > Articles > Expression of H9N2 Subtype Avian Influenza Virus HA Gene and Preparation of its Mono-specific Serum China Animal Husbandry & Veterinary Medicine 2015,42 (11) 2888-2894

H9N2亚型禽流感病毒HA基因的表达及其单因子血清的制备

作  者：
陈俊霞;孙鹏;许书珍;李思菲;苏帅;赵鹏;崔治中
单  位：
山东农业大学动物科技学院
关键词：
H9N2亚型禽流感;HA基因;表达;间接免疫荧光试验(IFA);单因子血清
摘  要：
为了制备特异性识别H9N2亚型禽流感病毒(AIV)的单因子血清,本试验提取H9N2亚型AIV RNA,RT-PCR后,分别扩增上段HA1、中段HA2和下段HA3 3段基因。将他们插入原核表达载体pET-32a(+)中,转化BL21(Rosetta)菌株中表达。将表达的蛋白常规免疫昆明白小鼠,以制备抗血清。结果显示,成功获得3段重组融合蛋白,且均具有良好的免疫原性。间接免疫荧光试验(IFA)结果显示,上段HA1、中段HA2制备的单因子血清均可与H9N2亚型AIV反应,而下段HA3则不能。重组马立克氏病病毒(MDV)MZC12 HA/NA同样证明HA1、HA2两段制备的单因子血清能识别HA基因的表达。本试验成功制备了识别H9N2亚型AIV HA的单因子血清,为H9N2亚型AIV的鉴别诊断及研究奠定了基础。
译  名：
Expression of H9N2 Subtype Avian Influenza Virus HA Gene and Preparation of its Mono-specific Serum
作  者：
CHEN Jun-xia;SUN Peng;XU Shu-zhen;LI Si-fei;SU Shuai;ZHAO Peng;CUI Zhi-zhong;College of Animal Science and Veterinary Medicine,Shandong Agricultural University;
关键词：
H9N2subtype avian influenza;;HAgene;;expression;;indirect immunofluorescence assay(IFA);;mono-specific serum
摘  要：
To prepare the mono-specific serum to diagnose H9N2 avian influenza virus(AIV),this test extracted H9N2 subtype AIV RNA and then amplified the upper HA1 gene,the middle HA2 gene and the lower HA3 gene by RT-PCR,respectively.Then they were inserted into expression vector pET-32a(+)and transformed into BL21(Rosetta)expression strain.The expressed proteins were used to immune Kunming White mice to prepare antiserum.Recombinant fusion proteins of HA1HA2 and HA3were obtained successfully and they showed good immunogenicity.Indirect immunofluorescence assay(IFA)showed that the two serums obtained by the upper HA1 and the middle HA2 could react with the H9N2 subtype AIV,while that of the lower HA3 could not.Recombinant Marek's disease virus(MDV)MZC12HA/NA also proved that the serums prepared by HA1 and HA2could recognize the expression of HA gene.The mono-specific serum of H9N2 subtype AIV was prepared successfully,which could lay the foundation for the diagnosis and research of H9N2 subtype AIV.

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