猪生殖—呼吸道综合征病毒(PRRSV)E蛋白基因真核表达质粒的构建

CONSTRUCTION OF A PLASMID EXPRESSING GENE ENCODING E.PROTEIN OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME(PRRS)VIRUS STRAIN CH-1A

新近发现的ＰＲＲＳＶ是单股ＲＮＡ病毒，属于不久前成立的动脉炎病毒科。其基因组包括８个开放阅读框架（ＯＲＦ），其中ＯＲＦ５编码糖基化的囊膜（Ｅ）蛋白，是病毒的主要免疫保护性抗原之一。为了研制ＰＲＲＳ基因疫苗，扩增并克隆了ＰＲＲＳＶ－１ａ株ＯＲＦ５，并将其亚克隆至真核表达载体ｐＣＲ３－Ｕｎｉ的ＣＭＶ启动子下游，构建成真核表达质粒。

Abstract Porcine reproductive and respiratory syndrome virus (PRRSV)is a newly recognized enveloped singlestrand RNA virus, belonging to a recently approved family, Arteriviridae. Its genome consists of 8 open reading frames (ORFs), with ORF5 encoding enveloped glycoprotein (E protein). The fragment encompassing entire ORF5 of PRRSV stain CH-la was amplified by reverse transcription and polymerase reaction(RT-PCR)and cloned into pUC18, and then subcloned into pCR3 Uni, an eucaryotic expressing vector. The constructed expressing plasmid pCR-ORF5 will be used for future gene vaccination