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Using SecM Arrest Sequence as a Tool to Isolate Ribosome Bound Polypeptides
Published on: June 19, 2012
Isolation, expression, and functional analysis of developmentally regulated plasma membrane polypeptide 1 (DREPP1) in
Cattarin Theerawitaya1, Nana Yamada-Kato2, Harminder Pal Singh3
1National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathum Thani, 12120, Thailand.
Abstract:
The plant specific DREPP proteins have been shown to bind Ca2+ and regulate the N-myristoylation signaling and microtubule polymerization in Arabidopsis thaliana. The information about DREPP proteins in other plants is, however, scarce. In the present study, we isolated the DREPP gene from a halophytic grass, Sporobolus virginicus, and tested whether the gene was involved in alkaline salt stress responses. The SvDREPP1 was cloned from S. virginicus by RACE methods. The isolated gene showed high homology to DREPP homologs from C4 grasses, Setaria italica, and Panicum hallii as well as rice (OsDREPP1). The encoded protein contained 202 amino acid residues. It was expressed in E. coli, and its biochemical properties were studied. It was observed that SvDREPP1 was not only Ca2+-binding protein, but also bind to calmodulin and microtubules. The SvDREPP1 mRNA expression in plants grown under alkaline salt stress was upregulated by 3.5 times over the control in leaf tissues after 48-h treatment, whereas it was increased for 6.0 times in the root tissues at 36 h. The data suggests the importance of SvDREPP1 in regulating alkali salt stress responses in the leaf tissues.
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