山羊痘病毒P32基因的PCR—RFLP分析

根据羊痘病毒基因组设计引物,建立了检测羊痘病毒P32基因的PCR方法,并扩增了疫苗毒株、标准毒株、贵州分离株的P32基因;应用生物学软件对山羊痘和绵羊痘病毒P32基因的酶切位点进行分析,选择合适的限制性内切酶对P32基因进行酶切.结果表明,以SDS-蛋白酶K法提取的病毒DNA在含量和纯度上均高于Trizol法,更有利于PCR扩增;所建立的PCR方法对细胞感染物及山羊痘疹样本均能扩增出特异性的目的DNA条带;软件分析选择限制性内切酶Hinf I对羊痘病毒P32基因的PCR产物进行酶切鉴定,可以有效区分山羊痘病毒和绵羊痘病毒.

Analysis on P32 Gene of Goatpox Virus by PCR-RFLP

The PCR method of detecting Capripoxvirus was set up by designing the primers according to the goatpox virus gene sequences published in the Genbank database.The P32 genes of vaccine strain Y,standard strain B,isolated strain LD and QL can be amplified by the PCR method.The PCR products were then digested by the restriction enzyme Hinf I selected by the biological software.The results showed that content and purity of viral DNA extracted by SDS-proteinase K were better than that by Trizol,which is beneficial to PCR amplification.The goatpox virus and sheeppox virus can be discriminated according to digesting their P32 gene by the restriction enzyme Hinf I.