%0 Journal Article
%T Cloning of full-length genes of H5N1 subtype Avian influenza virus strain A/duck/Shandong/093/2004 and analysis of the sequences<br>H5N1亚型禽流感病毒A/duck/Shandong/093/2004株的全基因克隆及序列分析
%A LONG Jin-xue
%A WANG Qu-zhi
%A LU Jian-hong
%A LIU Yu-liang
%A LIU Xiu-fan
%A <br>龙进学
%A 王曲直
%A 卢建红
%A 刘玉良
%A 刘秀梵
%J 微生物学报
%D 2005
%I 
%X The eight full-length genes, including the 5' and 3' ends of H5N1 subtype Avian influenza virus (A/duck/ Shandong/093/2004) were amplified by using the universal primers and H5 specific primers. The method used for the amplification of Avian influenza virus's full-length sequence was more easily and rapidly than that of rapid amplification of cDNA ends assay (RACE). The amplified segments were cloned into the T vector PCR 2.1, respectively. Three to five positive clones of each gene were sequenced and the same two sequencing results of the full-length genes were obtained. The phylogenetic analysis results showed that all the eight segments of the A/duck/Shandong/093/2004 were different from the A/Quail/Hongkong/G1/97 and A/Chicken/Beijing/1/94, but showed highly similarity (99% and above) to that of four H5N1 strains, which were isolated in 2002 in duck. It revealed that this strain was resulted from re-assortment of H5N1 rather than H9N2. The NA sequence of A/D/SD/04 was analyzed and the result demonstrated that there are 20 amino acids missing in 48 - 68 sites, however, there was no residue lost in NS gene in 263 to 277 sites. The motif of HA cleavage site is PQRERRRKKR/G, which is the characteristic of HPAIV. The 226 amino acid residue was Met (M), which can react with both Aalpha-2, 3Gal and SAalpha-2, 6Gal receptor. And the 627 residue of PB2 was Glutamic acid (E). The result mentioned above confirmed that H5N1 subtype AIV has multiple determinants in its virulence. A/D/SD/04 is the mid-strain evolving from HPAIV to a virulent strain of mammal.
%K H5N1 Subtype influenza virus
%K Universal primer
%K Full-length genes
%K Sequence analysis<br>H5N1亚型禽流感病毒，通用引物，全基因组序列，序列分析
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=32A04EEF3C176CFD&yid=2DD7160C83D0ACED&vid=94E7F66E6C42FA23&iid=94C357A881DFC066&sid=DC2727B6AC017B3F&eid=A7379F6713A46835&journal_id=0001-6209&journal_name=微生物学报&referenced_num=13&reference_num=10