A亚群禽白血病病毒QC6281株的分离与gp85基因序列分析

本试验通过病理剖检、接种DF-1细胞、ELISA及RT-PCR检测,从北京某种鸡场的疑似禽白血病的父母代蛋鸡中分离到1株A亚群禽白血病病毒,命名为QC6281.并根据GenBank中已发表序列设计引物,扩增env基因片段并将其克隆到pMD19-T-simple 载体中,扩增的env基因片段大小为1 239 bp,其中包括完整的gp 85基因,将gp 85基因亚克隆到pMD19-T-simple载体中并测序,gp 85基因大小为1 032 bp,编码344个氨基酸.将env基因片段和推导的氨基酸序列与GenBank中9株ALV的env基因相应序列做同源性分析并制作进化树图谱.发现QC6281与已发表的A亚群禽白血病病毒株该区域核苷酸同源性在83.8%～98%,氨基酸同源性在86.9%～98.5%.其中与Myeloblastosis-associated virus type 1(L10922.1)同源性最高,核苷酸同源性为98%,氨基酸同源性为98.5%;且与Myeloblastosis-associated virus type 1(L10922.1)亲缘关系最近.本研究为该毒株的生物学特性以及致病性研究打下基础.

Isolation of subgroup A avian leukosis virus QC6281 and partial sequence comparison

In this study,subgroup A avian leukosis virus was isolated by pathological anatomy,DF-1 cell culture,ELISA,RT-PCR from parent egg chickens and designated as QC6281.According to the sequences of env that was published on GeneBank,a pair of primers was designed.Partial env gene of QC6281 was amplified and cloned into pMD19-T-simple vector and sequenced.The nucleotide sequence of env gene was 1 239 bp.A open reading frame of gp85 gene was included,which was subcloned to pMD19-T-simple vector and sequenced.The ...