A Step-by-Step Protocol for Classifying AOX Proteins in Flowering Plants.

The potential of alternative oxidase (AOX) genes to develop functional markers for plant breeding programs has been emphasized. In this sense, it is essential to have a reliable classification system, which could aid in the selection of candidate AOX genes from different species. In the case of angiosperms AOX, a robust classification system is required because this enzyme is encoded by variable gene numbers (1-6 genes) with variable AOX subfamilies and subtypes. Thus, in this protocol, we present a detailed guideline to application of a classification scheme of AOX based on specific amino acids and phylogeny. We believe that this classification protocol provides an easier and practical way of classifying new angiosperm AOX genes besides that it can help to standardize AOX gene names used in AOX research community.