鸭源流感病毒分离株表面蛋白基因的克隆及序列分析

The Sequence Analysis the Surface Glycoprotein Genes of a Duck Origin Influenza virus Isolate

从湖南分离得到一株H5N1亚型禽流感病毒（AIV），首先设计合成两对HA和NA基因特异性引物，采用两步法RT-PCR，对鸭流感病毒株A／Duck／Hunan Wugang／2／2004（H5N1）（简称DK／HNWG／2／04）的表面蛋白基因进行序列测定，并与国内外已经发表的H5N1亚型毒株表面蛋白基因进行序列分析和比较。结果表明，HA和NA基因全长分别约为1．7kb和1．4kb，分离株与14个参考毒株HA基因的同源性为95．9％～98．0％，NA基因的同源性为87．0％～98．4％。根据HA基因核苷酸序列推导HA裂解住点氨基酸，发现分离株的裂解位点包含多个碱性氨基酸，符合高致病性禽流感的特征。

In this study, an avian influenza viruses （AIV）was isolated from Hunan province, and it was identified to be H5N1. According to the published sequence of H5N1 HA and NA genes, two pairs of specific primers were synthesized. The whole genetic encoding of the isolated strain of HA and NA genes were about 1.7 kb and 1.4 kb nucleotide with a single opening frame. The homology analysis showed that the homology of HA genes between the isolate and twelve reference viruses was 95.9 ～98.0 , the homology of NA genes was 87.0%～98.4%. The deduced analysis of HA genes splitting site suggested that the nucleotides contain characteristic sequence of splitting site of high pathogenic AIV strain.