鹅源副黏病毒NA-1株全长cDNA克隆的构建及鉴定

Construction and identification of the full-lenth cDNA clone of GPMV NA-1 strain

应用cRACE法扩增鹅源副黏病毒NA-1株cDNA 5′末端。参照Peter等的方法设计引物,扩增鹅源副黏病毒NA-1株cDNA的3′末端。根据GenBank上发表的鹅源副黏病毒序列设计6对引物,应用RT-PCR方法分6段扩增此病毒结构基因序列,并将其连接完成后与连接有鹅源副黏病毒NA-1株末端序列的转录载体连接,构建得到全长cDNA克隆。鹅源副黏病毒NA-1株全长cDNA克隆成功构建,为下一步病毒的拯救奠定坚实的基础。

Goose-host paramyxovirus strain NA-1 eDNA 5＇-terminal end were determined by cRACE; The primers were designed described by Peter etc to determine the goose-host paramyxovirus strain NA-1 cDNA 3＇-terminal end. Six pairs primer were designed according to the viral sequence published on GenBank to amplify this virus struc-tural gene sequence by RT-PCR. The structural gene sequence were connected with transcription vector which con-nected with the 3＇-and 5＇-terminal ends and the full-lenth eDNA clone was constructed correctly proved by sequencing. Goose-host paramyxovirus strain NA-1 full-lenth eDNA elong was constructed successfully, which laid a good foundation for the further rescued research.