摘要
运用RT-PCR技术扩增了禽流感病毒A／Chicken／Jiangsu／JS-1/2002(H9N2)完整的血凝素 (HA)基因,并克隆到pGEM-T载体中,进一步进行了序列测定。序列测定结果已经登陆 GenBank,登陆号为AY364228。所扩增的HA基因长度为1683核苷酸,共编码560个氨基酸,其中 信号肽长度为18aa,HA1为319aa,HA2为223aa。HA蛋白裂解位点的氨基酸组成为RSSR↓GLF, 不舍连续的碱性氨基酸,具有低致病性AⅣ HA基因裂解位点的序列特征。通过构建一个包含 18株H9N2亚型AIV的HA基因遗传进化树,发现所有的18个毒株共可分为欧亚谱系和北美谱 系两个谱系,而本分离株在分类地位上国内另外的三个分离株同属于欧亚谱系,在遗传进化上非 常接近,表明它们可能具有共同的起源。
RT-PCR were employed to amplify the cDNA of hemagglutinin（HA） gene of A/Chicken/Jiangsu/JS-1/ 2002（H9N2） and the cDNA were cloned into pGEM^R -T vector and sequenced. The sequencing result has logging in GenBank, the accession number was AY364228 .The result revealed that the HA genes of JS-1 had a complete open reading frame （ORF） and composed of 1683 nucleotides, coding for 560 amino acids. The deduced amino acid sequence at the cleavage site of the HA protein was RSSR ↓ GLF, it was match to the characteristic of low virulent strain. A phylogenetic tree based on the nucleotide sequences of the HA gene revealed all the 18 strains were divided into two lineages, namely Eurasian lineage and North America lineage, the isolate JS-1 was similar to the other three strains isolated from China, included A/Chicken/Beijing/1/94, A/Chicken/Guangdong/SS/94 and A/Chicken/ Guangxi/KMl/99, belonged to the Eurasian lineage.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2005年第B04期273-277,共5页
China Biotechnology
