禽流感病毒H5N1血凝素蛋白的重组牛痘病毒表达

目的研究H5 I151F和H5 I151F＋A134V＋E186D两种氨基酸变异对血凝素蛋白（HA）亲人受体结合的影响，并获得该HA。方法构建载体pRB21’，共感染／转染vp37^-牛痘病毒vRB12和pRB21’，基因同源重组牛痘病毒表达HA，Western免疫印迹法鉴定。结果获得了克隆有H5HA全长基因片段的载体pRB21’，构建了2种重组牛痘病毒re-VV H5 I151F和re-VV H5 I151F＋A134V＋E186D，且能在被感染细胞膜表达这两种HA。结论首次通过构建重组牛痘病毒成功表达了禽流感病毒H5N1的H5HA，为进一步研究禽流感病毒人传人的可能性奠定了基础。

Avian influenza virus H5N1 hemagglutinin proteins expressed by recombinant vaccinia viruses

Objective To require hemagglutinin （HA） proteins for the study on the relationships between two kinds of amino acid alterations （H5 I151F, H5 I151F ＋ A134V ＋ E186D） and human receptor binding characteristics. Methods Vector pRB21＇ was constructed, vp37^- vaceinia virus vRB12 and pRB21＇ were co-infected and transfected. Recombinant vaccinia viruses （re-VV） were generated through homologous recombination, expressing HA proteins, which were identified with Western Blotting. Results The vector pRB21 ＇ within full length of H5 HA segments were obtained. Two kinds of HA proteins （re-VV H5 I151F, re-W H5 I151F＋ A134V＋ E186D） were expressed by recombinant vaccinia viruses. Conclusion For the first time recombinant vaccinia viruses were generated to express hemagglutinin of the avian influenza virus subtype H5N1, which pave the way for further studying the possibilities of human to human transmission caused by avian influenza viruses.