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目的研究H5I151F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-VVH5 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。
Objective To study the effect of two amino acid mutations of H5I151F and H5I151F + A134V + E186D on the binding of hemagglutinin protein (HA) to human receptors and to obtain the HA. Methods Construction of vector pRB21 ’, co-infection / transfection of vp37-vaccinia virus vRB12 and pRB21’, homologous recombination vaccinia virus expressed HA and Western blotting. Results The vector pRB21 ’cloned with full length H5HA gene was obtained. Two kinds of recombinant vaccinia virus re-VVH5 I151F and re-VV H5 I151F + A134V + E186D were constructed and expressed both in the infected cell membrane. Conclusions The H5HA of the bird flu virus H5N1 was successfully expressed for the first time by constructing a recombinant vaccinia virus, which lays a foundation for further research on the possibility of human transmission of avian influenza virus.