鹅Ⅰ型禽副粘病毒融合蛋白基因3′端cDNA片段的分子克隆

Molecular Cloning of the 3′ end cDNA Fragments of the Gene Encoding the Fusion Protein of an Avian Paramyxovirus Type 1 Isolated from Goose

参考禽副粘病毒的融合蛋白基因 (F基固 )cDNA序列 ,设计并合成了 1对引物 ,以分离到对鹅有致病力的Ⅰ型禽副粘病毒GPMV/QY97 1株的核酸 (RNA)为模板 ,用逆转录聚合酶链反应 (RT PCR)对其融合蛋白基因 3′端进行扩增 ,获得了预计的 884bp左右的片段。将该片段克隆进 pGEM TEasy质粒载体 ,获得重组质粒T GPMVF3′ ,采用限制性内切酶和PCR方法对该重组质粒进行鉴定 ,证明所克隆的片段即为目的基因片段 。

According to the cDNA sequence of the gene (Fgene) encoding the fusion protein of an avian paramyxovirus, one pair of primers have been designed and synthesized. Using the genome RNA of an avian paramyxovirus type 1( APMV 1) strain GPMV/QY97 1 isolated from an affected goose as template, the 3′ end cDNA fragments of F protein gene of strain GPMV/QY97 1 were amplified by reverse transcription polymerase china reaction (RT PCR) and cloned into plasmid pGEM T Easy vector, the recombinant T GPMVF3′ was obtained. The recombinant was identified by a method of restriction endonuclease digestion and PCR, proving the cloned fragments was in the correct position. It may lay the foundations for further molecular biological study of APMV 1.