摘要
从鸡胚尿囊液中提取H9N2亚型禽流感病毒的 RNA,根据已发表的 A型流感病毒(AIV)的核苷酸序列 ,设计一对特异引物 ,采用反转录-聚合酶链式反应 (RT-PCR)成功地扩增了AIV的M基因.将M基因的cDNA克隆后进行了序列测定,测序结果表明所扩增的1 192 nt片段包含了完整的M基因的两个开放阅读框.核苷酸序列比较分析结果表明:该毒株与A/Hong Kong/1073/99(H9N2)、A/Duck/Hong Kong/380.5/2001(H5N1)、A/Duck/NY/191255-79/02(H5N2)相比 ,核苷酸序列的同源性在92.2 %~96.1 %之间,其相对应的氨基酸序列的同源性在91.7 %~96.3 %之间.
The Matrix(M) through encoding cDNA of H9N2 subtype avain influenza virus(AIV) was amplified and cloned. The virus RNA was directly extracted and purified from chicken embryo allantoic fluid. According to published gene sequence, a pair of specific primers to the M gene was designed and synthesized. After that, the M cDNA was amplified by reverse trascription--polymerase chain reaction (RT--PCR). Then the complete M gene was sequenced. The results showed that the cloned 1 192 nt fragment covered the whole M gene including the completely two open reading frame. When comparing with A/Hong Kong/1073/99 (HgN2), A/Duck/Hong Kong/380. 5/2001 ( HSN1 ) and A/Duck/NY/191255 -- 79/02(H5N2), the homology of nucleotide sequence ranged from 92.2 % to 96.1 % and homology of deduced amino acid was from 91.7 % to 96.3% .
出处
《甘肃农业大学学报》
CAS
CSCD
2005年第5期579-582,共4页
Journal of Gansu Agricultural University
基金
农业部畜禽病毒学重点开放实验室资助课题(2002-06)