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Expression, purification
and kinase activity analysis of maize ZmSPK1, a member of
plant SnRK2 subfamily
Huawen Zou1*,
Guohui Ma1,2, Cuihua Li1 and Xiaohai
Tian1
1College
of Agriculture, Yangtze University, Jingzhou 434025, China.
2China
National Hybrid Rice Research and Development Center,
Changsha 410125, China.
*Corresponding author.
E-mail:
zouhuawen73@hotmail.com.
Accepted 19 May, 2009 |
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Kinase
activity is essential for a protein kinase to perform its
biological function. In previous study we have cloned a
novel plant SnRK2 subfamily gene from maize and named it as
ZmSPK1. In this study the cDNA of ZmSPK1 with
dHA-His6 tag was amplified by PCR and was subcloned into the
yeast expression vector p426GAL1. The constructed plasmid
named as p426GAL1-SPK1-dHA-His6 was transformed into
Saccharomyces cerevisiae.
Under the induction of galactose the recombinant protein was
expressed. The purified recombinant protein showed a single
specific band by the analysis of the western-blot. In an
in vitro kinase assay, the purified recombinant protein
showed phosphorylation and autophosphorylation activity.
This showed that ZmSPK1 encodes a functional protein kinase.
Key
words:
Zea mays L., ZmSPK1, kinase activity. |
