Purification and characterization of the recombinant Bambusa oldhamii Monocot-Specific Protein-41
Date Issued
2014
Date
2014
Author(s)
Lee, Pei-Ju
Abstract
BoMSP41 (Monocot-specific protein-41 in Bambusa oldhamii), an unknown function gene, is highly up-regulated in the internode-containing region of rapidly elongating bamboo shoots. The protein encoded by this gene was suggested to be an intrinsically disordered protein (IDP) and might be translocated into chloroplasts. To understand the characteristics of BoMSP41, the His-tagged recombinant BoMSP41 proteins produced in E. coli were purified. The recombinant BoMSP41 had several unusual properties including a lower electrophoretic mobility on SDS-PAGE, overestimated molecular weight with gel filtration and high thermal stability, which are generally observed in many IDPs. Analysis of the purified recombinant BoMSP41 by circular dichroism (CD) spectroscopy suggested that there were 8.6% alpha-helix, 37.0% beta-sheet, 12.9% beta-turn and 41.5% random coil in BoMSP41. The results showed that BoMSP4 contained intrinsically disordered regions as previously predicted. The purified recombinant BoMSP41 proteins were used as antigen to raise polyclonal antibodies. Western analyses of proteins extracted from developing bamboo shoots with the polyclonal antibodies showed that the transcripts of BoMSP41 gene could be translated into proteins in planta and BoMSP41 might not be present in chloroplasts and nuclei. To perform its function, BoMSP41 might require binding partners to induce the transition of its disorder region to ordered conformation.
Subjects
bamboo
monocot-specific protein
recombinant protein
inclusion bodies
purification
circular dichroism
intrinsically disordered protein
subcellular localization
Type
thesis