Dupla difusäo radial na análise genética das lipoxigenases L1 e L2 em soja / Double gel immunodiffusion in the genetic analysis of lipoxygenases L1 and L2 from soybeans

ABSTRACT

In the effort to genetically manipulate soybean seeds aiming to improve flavor quality toward human consuption, lipoxygenase null alleles are being transferred to a brazilian commercial variety. Immunochemical approach has been taken to perform analysus of hundred of seeds derived from the breeding program. For that reason lipoxygenases L1 and L2 were puridied to hemogeneity by gel filtration and affinity chromatography. Policlonal antibodies were prepared in rabbits and purified further by affinity chromatogrphy. By using the purified anti-L1 and anti-L2 an Ouchterlony double immunodifusion method has been developed in our lab to detect the presence/absence of the isozymes simultaneously in soybean seeds. The method proved to be highly specific and efficient enought to perform single non destructive seed analysis: