Preparation Of Monoclonal And Polyclonal Antibodies Against Maize Resistance Protein ZmWAK And ZmTrxh

The two main diseases in corn production are Maize smut and dwarf mosaic disease,which affect almost all maize production areas in the world,and cause huge loss of quantity and quality.At present,the most economical and effective measure is to use corn's own resistance sources to select and breed superior resistant varieties of corn,which is to control the occurrence of smut and dwarf mosaic disease in maize.Therefore,it is vital to establish a fast and simple method for the detection of disease-resistance genes for the breeder's disease-resistant breeding of corn.In this study,maize anti-smut gene Zm WAK and anti-dwarf mosaic gene ZmTrxh were research objects,and colloidal gold immunochromatographic technique was used.These developed a method to detect maize disease resistance genes rapidly.The two kinds of genes expressed in Escherichia coli were purified effectively and were used to immunize mice and rabbits.Monoclonal antibodies against recombinant disease-resisting proteins were produced by hybridoma cell technique,and rabbit-derived polyclonal antibodies were also obtained.It offers a foundation for the detection of maize disease-resistance protein.The main results of this study are as follows:(1)The acquisition of Zm WAK and anti-dwarf mosaic gene ZmTrxh from maize and the construction of prokaryotic expression vector.(2)The prokaryotic expression,identification and purification conditions of maize recombinant disease-resistant protein.(3)BALB/c mice and New Zealand white rabbits were immunized with purified recombinant disease resistance protein to obtain the murine and rabbit-derived polyclonal antiserum against Zm WAK protein and ZmTrxh protein,and the immunological result indicated that the protein has good immunogenicity.(4)Cell fusion of mouse spleen cells and myeloma cells was performed by cell fusion technique and screened by competitive ELISA to obtain positive hybridoma cells.After subcloning,positive hybridoma cell lines that secrete recombinant Zm WAK protein with good immunological properties were obtained and designated respectively as 4C3-G8,5F7-E2,5F7-H3,and 7F4-C5.