Agriculture Reference
In-Depth Information
brassinolide, the two pea brassinosteroid C-6 oxidases (CYP85A1 and
CYP85A6) convert 6-deoxocastasterone to castasterone by C-6 oxidation
(Jager et al., 2007).
S
HORT
-C
HAIN
A
LCOHOL
D
EHYDROGENASE
-L
IKE
P
ROTEIN
(SAD)
The SAD gene family in pea has at least three proteins (SAD-A, SAD-B,
and SAD -C). Expression of SAD-A and SAD-C in Escherichia coli or Pichia
pastoris yields full-length soluble proteins. Tetrameric SAD-C protein is a
dimer of dimers. Following brief exposure to ultraviolet-B radiation SAD
genes are transiently expressed whereas after prolonged UV-B irradiation
mRNA accumulation results in formation of SAD protein in leaves and stems
(Scherbak et al., 2009).
F
ATTY
A
CID
A
LPHA
-D
IOXYGENASE
alpha-Dioxygenases catalyze the first step of the alpha-oxidation pathway
of long-chain fatty acids in plants.They are expressed in plants in response to
biotic and abiotic stress. During germination, alpha-dioxygenase mRNA
accumulates in the cotyledons and the embryonic axis of pea seeds. In
developing pea plants, the transcript is detected in roots. The accumulation of
alpha-dioxygenase protein parallels transcript accumulation in that it is
abundant in germinating as well as young plant tissue, and correlates with loss
of mRNA during plant maturation (Meisner et al., 2009).
G
IBBERELLIN
D
IOXYGENASES
Pea gibberellin-oxidase was expressed as a glutathione s-transferase
fusion and purified with a yield of 0.3 mg/g cells by affinity chromatography,
glutathione s-transferase removal and gel filtration. It displays a Km of 0.024
microM and a Vmax of 4.4 pkat/mg toward [1beta, 2beta, 3beta-3H3]GA20.
Fusion tags are needed to stabilize and solubilize PsGA2OX1 during
E. coli
expression.
