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H5N1亚型禽流感病毒神经氨酸酶基因的表达及纯化
引用本文:张改平,李清州,郝惠芳,周玲,王丽,杨苏珍,罗俊,冯丽丽.H5N1亚型禽流感病毒神经氨酸酶基因的表达及纯化[J].华北农学报,2008,23(5).
作者姓名:张改平  李清州  郝惠芳  周玲  王丽  杨苏珍  罗俊  冯丽丽
作者单位:1. 河南省动物免疫学重点实验室,河南,郑州,450002
2. 河南省动物免疫学重点实验室,河南,郑州,450002;河南农业大学,牧医工程学院,河南,郑州,450002
3. 河南农业大学,牧医工程学院,河南,郑州,450002
基金项目:国家"973"计划项目
摘    要:参考GenBank上发表的H5亚型禽流感病毒(AIV)神经氨酸酶(NA)基因序列设计引物,PCR扩增NA基因,再将其克隆到原核表达载体pET-32a中,用E.coli BL21(DE3)原核表达,SDS-PAGE和Western-blot对表达蛋白进行鉴定.Western-blot结果表明,表达产物具有免疫学活性,蛋白的分子量约为61 kDa,位于包涵体中.包涵体经变性、复性处理,表达蛋白能与H5亚型AIV阳性血清发生特异性反应,具有良好的抗原性.ELISA检测结果表明,用此纯化蛋白作为包被抗原检测H5N1亚型AIV神经氨酸酶抗体具有良好的灵敏性.

关 键 词:禽流感病毒  H5亚型  神经氨酸酶(NA)  原核表达

Expression of NA Gene of Avian Influenza Virus Subtype H5N1 in Escherichia coli and Purification of the Expressed Product
ZHANG Gai-ping,LI Qing-zhou,HAO Hui-fang,ZHOU Ling,WANG Li,YANG Su-zhen,LUO Jun,FENG Li-li.Expression of NA Gene of Avian Influenza Virus Subtype H5N1 in Escherichia coli and Purification of the Expressed Product[J].Acta Agriculturae Boreali-Sinica,2008,23(5).
Authors:ZHANG Gai-ping  LI Qing-zhou  HAO Hui-fang  ZHOU Ling  WANG Li  YANG Su-zhen  LUO Jun  FENG Li-li
Institution:ZHANG Gai-ping1,LI Qing-zhou1,2,HAO Hui-fang1,ZHOU Ling1,WANG Li1,YANG Su-zhen1,LUO Jun1,FENG Li-li2 (1.Henan Key Laboratory for Animal Immunology,Zhengzhou 450002,China,2.Engineering College of Animal , Vetrinary Science,Henan Agricultural University,China)
Abstract:The neuraminidase(NA)gene of Avian intluenza virus(AIV)subtype H5 was amplified by PCR according to the sequence published on GenBank.It was cloned into the expression vector PET-32a.The protein encoded by this gene was expressed in Escherichia coli strain BL21(DE3)and was characterized by SDS-PAGE and western blotting analysis as a 61 kDa protein with immunogenicity.The fusion protein was present primarily in inclusion bodies and was purified via denaturation and renenaturation.The results indicated that t...
Keywords:Avian influenza virus  Subtype H5  Neuraminidase(NA)  Prokaryotic expression  
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