Preparation and Application of Monoclonal Antibody to Sugarcane (Saccharum L. Spp. Hybrids) Sop5cs Protein

Abstract

△ 1 -pyrroline-5-carboxylate synthase (P5CS) is a key enzyme in proline biosynthesis. In this study, the nucleotide sequence of sugarcane P5CS (SoP5CS) gene was obtained from sugarcane. Then the ORF of SoP5CS linked with pET-30a vector and the SoP5CS protein was expressed successfully in E. coli by the induction of 1.0 mM IPTG at 28 °C for 6 h. The prokaryotic expression protein was expressed as inclusion bodies (IBs). It was purified by Ni-NTA Resin firstly and then refolded, dialyzed and further purified by gel filtration. The monoclonal antibody of SoP5CS was prepared using mice. The expression of SoP5CS gene and the corresponding protein and physiological parameters associated with drought resistance were detected in sugarcane leaves under different drought stresses, they worked together to regulate the drought resistance process of sugarcane. This is the first report on the preparation of P5CS monoclonal antibody and the expression of the antibody in sugarcane, which would provide new methods to study the function of SoP5CS response to various abiotic stresses and other aspects.