传染性支气管炎病毒Holte株S1基因的克隆及鉴定

Cloning and Identification of S1 Gene of Avian Infectious Bronchitis Virus Strain Holte

为给鸡传染性支气管炎（ＩＢ）基因工程疫苗的研制提供基因储备，通过ＲＴ－ＰＣＲ扩增出ＩＢＶＨｏｌｔｅ株Ｓ１基因ｃＤＮＡ，其长度与理论值１７６２ｂｐ相符；Ｓ１基因内部位点经ＨａｅⅢ酶切所产生的片段，亦与理论值５４９、３４３、３２４、１９１、１８０、１７１ｂｐ６个片段大小一致。将Ｓ１基因ｃＤＮＡ克隆入ｐＵＣ１８，并对Ｓ１基因两翼进行了序列分析。结果表明，所克隆的Ｓ１基因与ＧｅｎＢａｎｋ中收录的ＩＢＶＨｏｌｔｅ株Ｓ１基因完全相符。

The cDNA (1 762 bp) of S1 gene of IBV strain Holte was amplified by RT PCR, and the amplified product cloned into pUC18 to provide the gene reserves for development of the genic engineering vaccine. The cloned cDNA was sequenced and digested with HaeⅢ and six fragments of 549 bp, 343 bp, 324 bp, 191 bp, 180 bp and 171 bp obtained. The results show that the S1 gene cDNA of IBV strain Holte is identical with that reported by Genbank.