鸡传染性支气管炎北京地方株S_1基因的克隆与鉴定

Cloning and identification of S 1 gene of infectious bronchitis virus Beijing strains

利用反转录套式聚合酶链反应（ＲＴｎｅｓｔｅｄＰＣＲ）技术从北京地区３株鸡传染性支气管炎病毒（ＩＢＶ）分离株（ＢＪ１，ＢＪ２，ＢＪ３）中扩增出１０５４ｂｐＳ１基因高变区。利用限制性内切酶ＳｉｎⅠ和ＰｓｔⅠ的酶切位点分析图谱进一步证实了ＲＴｎｅｓｔｅｄＰＣＲ的特异性。将３段Ｓ１基因分别克隆到ｐＵＣ１９质粒中，获得重组质粒ｐＵＣＩＢＶＳ１ＢＪ１，ｐＵＣＩＢＶＳ１ＢＪ２和ｐＵＣＩＢＶＳ１ＢＪ３。

Using a reverse transcription nested polymerase chain reaction (RT nested PCR), the 1054 bp long S 1 gene hypervariable regions of 3 strains of avian infectious bronchitis viruses (IBV, BJ 1, BJ 2, BJ 3) isolated from Beijing areas were amplified successfully. By means of the map of restriction enzymes SinⅠ and PstⅠ, the specificity of the RT nested PCR products were proved. The recombinant plasmids pUC IBV S 1-BJ 1, pUC IBV S 1-BJ 2 and pUC IBV S 1-BJ 3 were obtained by cloning the three S 1 genes into plasmids pUC 19.