PREPARATION OF ANTISERUM TO RECOMBINANT COAT PROTEIN FOR DETECTING STRAWBERRY MILD YELLOW EDGE VIRUS

Abstract

Strawberry mild yellow edge virus (SMYEV) is one of the most economically important viral pathogens infecting strawberries worldwide. Immunodiagnostics is a rapid and sensitive method for detecting viruses. Preparation of antiserum against recombinant coat protein (CP) has been used for some viruses. In this paper, the full length CP gene of SMYEV, which consisted of 729 bp and encoded 242 amino acid residues, was isolated from the SMYEV isolate SY05, and then cloned into the prokaryotic expression vector pGEX-6P-1. The recombinant prokaryotic expression vector carrying the SMYEV CP gene was introduced into Escherichia coli and the fused CP protein was expressed in E. coli cells treated with IPTG. The antiserum was produced after the rabbit was immunized with the purified CP protein. Two methods, ELISA and RT-PCR, were compared for detecting SMYEV, and the results show that ELISA was as not sensitive as RT-PCR.