新城疫病毒长春株NP、P、M蛋白基因的克隆与序列分析

Cloning and Sequence Analysis of NP,P,M Gene of Avirulent NDV Strain Changchun

参考新城疫病毒 (NDV)相关毒株基因组核苷酸序列 ,设计了 3对特异性引物 ,应用 RT- PCR一次性扩增出NDV长春株 NP、P、M基因的氨基酸编码区 (ORF)。将 NP、P、M基因插入质粒 p Bluescript KS(- )后测序。序列分析表明 ,长春株 NP、P、M基因的 ORF长度分别为 1470、1188、10 95 bp,分别编码 490、396、36 5个氨基酸 ,与发表的13株 NDV毒株相关基因相比较 ,核苷酸序列同源性分别为 85 .7%～ 99.1%、82 .8%～ 96 .9%、84.3%～ 99.7% ,推导的氨基酸序列同源性分别为 92 .5 %～ 99.3%、84.3%～ 95 .9%、88.9%～ 99.1%。对 NP、P、M基因同源性比较和进化树分析结果表明 ,长春株与 L a Sota、B1等弱毒株之间具有很近的亲缘关系 。

Three pairs of specific oligonucleotide primers for amplifying the NP, P and M gene of NDV were designed according to the published relevant sequences of NDV, and the NP, P and M gene of an virulent NDV strain( Changchun) were amplified by RT-PCR, and sequenced subsequently. The ORF of the NP, P and M gene is sequenced to be 1 470 bp,1 188 bp,1 095 bp which encod 490,396 and 365 amino acids respectively. Compared NP, P and M gene of strain Changchun with the published NDV strains (i.g. La sota, B1 et al.), the homology of the nucleotide sequence is 85.7%-99.1%,82.8%-96.9% and 84.3%-99.7%,and the homology of the deduced amino acids sequence is 92.5%-99.3%,84.3%-95.9%,88.9%-99.1%.