Functional analysis of abscisic acid 8'-hydroxylase.

Akira Endo, Mitsuhiro Kimura, Naoto Kawakami, Eiji Nambara

Department of Cell & Systems Biology, University of Toronto, Toronto, ON, Canada.

Methods in molecular biology (Clifton, N.J.) 2011

filter terms:

Abscisic acid (ABA) plays an important role in the control of seed dormancy and germination. Identification of hormone metabolism genes from a particular plant species of interest is an essential step in hormone research. The function of these gene products is validated by biochemical analysis using heterologous expression systems, such as E. coli and yeast. ABA 8'-hydroxylase is a subfamily of P450 monooxygenases and is encoded by CYP707A genes. CYP707A catalyzes the committed step in the major ABA catabolic pathway. In this chapter, we describe the methods for RNA extraction from seeds, cloning the CYP707A cDNAs, protein expression in yeast, and biochemical analysis of their gene products.

Citation

Akira Endo, Mitsuhiro Kimura, Naoto Kawakami, Eiji Nambara. Functional analysis of abscisic acid 8'-hydroxylase. Methods in molecular biology (Clifton, N.J.). 2011;773:135-47