[Prokaryotic expression, antiserum preparation and cytochemical analysis of TaMlo3 in wheat]

Mlo gene in wheat is critical to determine wheat broad spectrum disease resistance against the pathogenic powdery mildew fungus. According to wheat TaMlo3 cDNA sequence, the two DNA fragments encoding the intracellular loop 2 and C-terminus of wheat Mlo protein were cloned respectively into the vector pET-30a, and then the fusion polypeptides were expressed in E. coli BL21. The recombinant proteins were purified by Ni2+ -NTA agarose column, and then were used to immunize the rabbits respectively. Two polyclonal antisera were obtained and they could be specifically bound on the extrahaustorial membrane and extrahaustorial matrix of the powdery mildew fungus by immuno-gold labeling.