番鸭呼肠孤病毒MW9710株S1基因片段的克隆及序列分析

参考GenBank番鸭呼肠孤病毒(muscovy duck reovirus,MDRV)S1基因序列设计合成一对引物,对番鸭呼肠孤病毒MW9710株S1基因进行RT-PCR扩增,并对PCR产物进行了克隆和测序.结果显示扩增产物为300bp,与预期的目的片段大小一致,经PCR、酶切反应鉴定后克隆到pGEM-Teasy载体中,核苷酸序列经BLAST软件分析表明:番鸭呼肠孤病毒MW9170株S1基因的目的片段与番鸭呼肠孤病毒法国89026株同源性为91.7%,与鸡关节炎病毒S2基因同源性为68.7%,结果提示番鸭呼肠孤病毒MW9710株与鸡关节炎病毒亲缘距离较远.

Cloning and sequence analysis of the muscovy duck reovirus MW9710 strain S1 gene fragment

A pair of primers were designed and synthesized according to the reported muscovy duck reovirus(MDRV)S1 gene in GenBank.We used it to amplify the S1 gene of the muscovy duck reovirus MW9710 strain by RT-PCR,then the PCR products were cloned and sequenced.The results showed that the RT-PCR products was 300bp consistent with prospective fragment.The target fragment was cloned to pGME-T easy vector by PCR and enzyme-cutting identification and was sequenced.The analysis of BLAST software suggested that the target sequence had 91.7 % homology between MDRV MW9710 strain and MDRV 89026 France strain,but 68.7 % homology between MDRV MW9710 strain and ARV.The results indicated that there was a significant difference between MDRV MW9710 strain and ARV.