H1N1猪流感广东株血凝素基因的克隆与序列分析  

Cloning and Sequence Analysis of Hemagglutinin Gene of the Swine Influenza Strain(H1N1)Isolated in Guangdong

作　　者：胡永浩[1] 姚学萍[1] 李海燕[2] 赵有淑[1] 杨焕良[2] 陈化兰[2] 

机构地区：[1]甘肃农业大学动物医学院,兰州730070 [2]中国农业科学院哈尔滨兽医研究所农业部动物流感重点开放实验室,哈尔滨150001

出　　处：《畜牧兽医学报》2005年第2期172-176,共5页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：农业部动物流感重点开放实验室开放课题资助项目

摘　　要：用RT- PCR方法扩增H1N1亚型猪流感病毒广东分离株 A/Swine/GuangDong/711/2001 HA基因,对其进行了克隆和测序。结果显示,HA基因全长 1 771 bp,共编码 579 个氨基酸。A/Swine/GuangDong/711/2001HA基因编码的氨基酸序列中有 8 个糖基化位点,5 个位于 HA1 基因的第 27、28、40、104 和 287 位点,3 个位于HA2基因的21、153和213位点。与H1N1亚型猪流感经典毒株比较后发现,血凝素糖基化位点并不是高度保守的。将A/Swine/GuangDong/711/2001与自GenBank读取的1918年人流感毒株 A/South Carolina/1/18、1991 年人流感毒株A/MD/12/91和1997年猪流感毒株 A/Swine/Wisconsin/238/97 等进行核苷酸同源性比较分析,A/Swine/GuangDong/711/2001与A/South Carolina/1/18、A/MD/12/91和 A/Swine/Wisconsin/238/97 等毒株的核苷酸同源性分别为 93. 9%、94. 7%和 93. 9%。从系统发生树来看, A/Swine/GuangDong/711/2001 与 A/SouthCarolina/1/18和A/Swine/Wisconsin/238/97的亲缘关系相近。The hemagglutinin(HA) gene of a strain of swine influenza virus (H1N1), A/Swine/ GuangDong /711/2001, was amplified by RT-PCR. The PCR products were sequenced after cloning into pMD18-T plasmid and the sequence was compared with the HA gene sequences from others reference strains of influenza virus in GenBank. The result indicated that 1 771 bp fragment covered the complete HA open reading frame, and encoded 579 amino acid residues. There were 8 glycosylation sites in the amino acid sequences of A/Swine/GuangDong/711/2001. The 5 glycosylation sites were positioned at 27, 28, 40, 104 and 287 of HA1 gene, others at 21,153 and 213 of HA2 gene. Compared with typical strains of swine influenza viruses(H1N1), the glycosylation sites were not conservative. Among A/Swine/GuangDong/711/2001 and A/South Carolina/1/18, A/MD/12/91 and A/Swine/Wisconsin/238/97, the homology of the nucleotides were 93.9,94.7 and 93.9 respectively. Phylogenetic analysis showed that A/Swine/GuangDong/711/2001 had a close relationship with A/South Carolina/1/18 and A/Swine/ Wisconsin/238/97.

关 键 词：猪流感病毒 HA基因 位点 毒株 广东株 l基因 序列分析 人流感 血凝素基因 亚型 

分 类 号：S858[农业科学—临床兽医学] S852[农业科学—兽医学]