Reference "Isolation of a gene encoding a glycosylated cytokinin oxidase from maize"

Reference ID

7146

Title

Isolation of a gene encoding a glycosylated cytokinin oxidase from maize

Source

Biochemical and biophysical research communications, 1999, vol. 255, pp. 328-333

Authors (4)

Morris-R-O	Bilyeu-K-D
Laskey-J-G	Cheikh-N-N

Abstract

The major cytokinin oxidase in immature maize kernels was purified to
homogeneity. Selected tryptic peptides were used to design degenerate
oligonucleotide primers for PCR isolation of a fragment of the oxidase gene.
Hybridization of the PCR fragment to a maize genomic library allowed isolation
of a full-length cytokinin oxidase gene (ckx1). The gene encodes a protein of
approximately 57 kDa that possesses a signal peptide, eight consensus N-
glycosylation sequences and a consensus FAD binding sequence. Expression of ckx1
in Pichia caused secretion of active glycosylated cytokinin oxidase that
contains a substrate-reducible FAD. The gene displays sequence homology with a
putative oxidoreductase from Arabidopsis thaliana and with the fas5 gene from
Rhodococcus fascians.