CAJ | 학술논문

目的建立能有效表达禽流感病毒A/Hubei/1/2010( H5N1)无头HA( headless HA)的杆状病毒系统.方法利用RT-PCR扩增获得headless HA基因,克隆至杆状病毒转移载体pFastBac1,命名为pFastBac1headless HA.将pFastBac1headless HA转化到DH10Bac感受态细胞,获得重组穿梭质粒Headless HA-Bac.利用Bac-to-Bac杆状病毒表达系统,转染Sf9昆虫细胞,获得重组杆状病毒,并进行Western Blot和红细胞凝集试验检测.结果 Headless HA在Sf9细胞中能有效表达,不能使红细胞发生凝集.结论本研究为利用headless HA表达蛋白研制广谱流感疫苗奠定了基础.
목적 건립능유효표체금류감병독A/Hubei/1/2010( H5N1)무두HA( headless HA)적간상병독계통.방법 이용RT-PCR확증획득headless HA기인,극륭지간상병독전이재체pFastBac1,명명위pFastBac1headless HA.장pFastBac1headless HA전화도DH10Bac감수태세포,획득중조천사질립Headless HA-Bac.이용Bac-to-Bac간상병독표체계통,전염Sf9곤충세포,획득중조간상병독,병진행Western Blot화홍세포응집시험검측.결과 Headless HA재Sf9세포중능유효표체,불능사홍세포발생응집.결론 본연구위이용headless HA표체단백연제엄보류감역묘전정료기출.
Objective To construct recombinant headless HA of avian influenza H5N1virus based on baculovirus expression system.Methods Headless HA gene of an avian H5N1virus,A/Hubei/1/2010 was cloned into pFastBac1vector.The recombinant shuttle vector,headless HA-Bac,was obtained by transforming pFastBac1headless HA into DH10bac cell and then the baculovirus bearing with headless HA gene,headlessHA-Bac,was achieved viaBac-to-Bac baculovirus expression system. The headless HA expression was detected in headless HA-Bac-infected Sf9 cells by Western Blot and HA assay.Results The headless HA has been efficiently expressed and displayed no reaction with turkey red blood cells.Conclusion The constructs provide fundamental work for future studies on universal influenza vaccine.