Plant Molecular Biology Reporter

, Volume 29, Issue 3, pp 739–744 | Cite as

Expression, Purification, and Secondary Structure Prediction of Pentatricopeptide Repeat Protein RF1A from Rice

Article
First Online:

Abstract

Pentatricopeptide repeat protein (PPR) proteins are putative RNA-binding proteins which are particularly prevalent in terrestrial plants. Previous research has reported the great difficulty in purifying soluble PPR proteins in Escherichia coli, therefore hindering further study of their functions. In this paper, we report the use of the pMAL ™ prokaryotic expression system to acquire a soluble expression of a PPR protein, RF1A from rice (Oryza sativa L.). After purification, we identified RF1A by ESI-TOF-MS/MS. We also made an estimation of its secondary structure using the circular dichroism spectroscopy. These results supported the bioinformatic prediction of helical-hairpin model about PPR proteins.

Keywords

RF1A Soluble expression Protein purification Circular dichroism 

Abbreviations

PPR

Pentatricopeptide repeat

DTT

Dithiothreitol

CD

Circular dichroism

SP

Sulfopropyl

CHT

Ceramic hydroxyapatite

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Notes

Acknowledgements

This work was supported by Chinese national 973 program (Grant number: 2007CB109005) and National Natural Science Foundation of China (30571144).

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  1. 1.Key Laboratory of MOE for Plant Developmental Biology, College of Life SciencesWuhan UniversityWuhanPeople’s Republic of China

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