畜牧兽医学报 ›› 2005, Vol. 36 ›› Issue (2): 172-176.doi:

• 研究报告 • 上一篇    下一篇

H1N1猪流感广东株血凝素基因的克隆与序列分析

胡永浩;姚学萍;李海燕;赵有淑;杨焕良;陈化兰   

  1. 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
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2005-02-25 发布日期:2005-02-25

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

HU Yong-hao;YAO Xue-ping; LI Hai-yan; ZHAO You-shu;YANG Huan-liang;CHEN Hua-lan   

  1. 1.College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070,China;2. Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2005-02-25 Published:2005-02-25

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/711/2001 HA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/South Carolina/1/18和A/Swine/Wisconsin/238/97的亲缘关系相近。

Abstract: 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.