禽呼肠病毒P10、P17非结构蛋白基因的克隆及序列分析

根据GenBank上的禽呼肠病毒(ARV)S1基因序列,设计并合成了一对跨越P10和P17非结构蛋白基因的特异性引物,对13个ARV毒株进行RT-PCR扩增、克隆及序列测定。结果显示,13个ARV毒株的P10蛋白基因ORF全长均为297bp,编码98个氨基酸;P17蛋白基因ORF全长为441bp,编码146个氨基酸。这13个ARV毒株P10、P17蛋白基因核苷酸同源性分别在96.6%~100%和95.2%~99.3%之间,推导的氨基酸同源性分别在98.2%~100%和91.9%~99.0%之间。将这13个ARV毒株与GenBank上其他正呼肠病毒毒株,包括番鸭株(DRV)和飞狐上分离到的内尔森海湾病毒(NelsonBayvirus,NBV)及两个澳洲分离株(ARM-1和SOM-4)进行同源性比较和遗传进化树分析,结果表明,呼肠病毒有地域和种类的差别。

Cloning and Sequnce Analysis of the P10 and P17 Nonstructural Protein Gene of Avian reovirus

A pair of specific primers were designed and synthesized according to S1 gene sequences of Avian reovirus from GenBank, which included the whole open reading frame (ORF) of P10 and P17 nonstructral protein genes.Both P10 and P17 protein genes of 13 ARV strains were amplified,cloned and sequenced. The whole ORF of P10 and P17 protein genes of those 13 ARV strains were 297bp and 441bp in length, respectively. And they were predicted to encode P10 and P17 nonstructral proteins of 98 and 146 amino acids, respectively. The percentages of nucleotide and amino acid sequence identities were determined. The nucleotide and amino acid sequence identities of P10 protein gene among those 13 ARV strains exhibited 96.6%-100% and 91.9%-99.0%,respectively. The identities of P17 protein gene showed 98.2%-100% and 95.2%-99.3%,respectively.Comparison of the nucleotide and predicted amino acid sequences and phylogenetic analysis between ARV and other orthoreoviruses(DRV,NBV,RAM-1,SOM-4) showed that Reovirus had the divergence among regions and species.