猪繁殖与呼吸综合征病毒GP5羧基端抗原表位的鉴定

Identification of an Antigenic Epitope Located in Carboxy-terminal of GP5 of Porcine Reproductive and Respiratory Syndrome Virus

为了鉴定和分析猪繁殖与呼吸综合征病毒(PRRSV)GP5羧基端的抗原表位,采用Goldkey软件分析PRRSVGP5羧基端抗原表位,经人工合成的方法获得编码GP5的部分基因,Eag 酶切后插入经Eag 线性化处理的噬菌体M13KEgIII克隆载体,转化至ER2738细胞,得到多株重组噬菌体,提取噬菌体的ssDNA进行PCR及序列鉴定,获得了展示GP5羧基端11个氨基酸的重组噬菌体,用PRRS阳性血清检测重组噬菌体,结果显示噬菌体展示的表位可被PRRSV感染血清所识别,从而证明GP5羧基末端的11个氨基酸是PRRSV的一个抗原表位。

In order to identify and analyse carboxy-terminal antigenic epitope of porcine reproductive and respiratory syndrome virus GP5, the GP5190 gene was synthesized and the fragments were digested by restriction endonuclease EagⅠ. The DNA fragments were ligated with vector, phage M13KE, which has been cut with EagⅠ. Then the recombinants were transfected into E.coli ER2738. The single strain DNAs of phages were extracted, and the insert was amplified by PCR and sequenced. The results showed that the recombinant phages contain the 11 amino acid epitope of PRRSV GP5, the ELISA result showed that this epitope could be recognized by antibody to PRRSV, indicating that the linear 11 amino acid is an antigenic epitope of porcine reproductive and respiratory syndrome virus.