Cloning and expression of Hsp22.4 gene from Chaetomium globosum

Abstract

A study was conducted on the molecular mechanism of small heat shock proteins (sHSPs) in Chaetomium globosum. Heat shock protein 22.4 (Hsp22.4) from C. globosum was cloned and expressed in Escherichia coli. BlastX analysis revealed that the Hsp22.4 gene from C. globosum shared the highest identity in amino acid sequence with a Hsp gene from Neurospora crassa, and the identity between them was 65%. The C. globosum Hsp22.4 gene was inserted into the expressive vector of pGEX-4T-2 and the recombinant plasmid named pGEX-HSP. E. coli BL21 transformed with pGEX-HSP plasmid was induced by IPTG, and the expressed proteins were analyzed with SDS-PAGE. A 50 kD protein was specially expressed in E. coli BL21, and the result was consistent with expectation, and showed that the Hsp22.4 gene had been expressed in E. coli. Our study has made a foundation for further studying the function of sHSPs protein.

Keywords

Chaetomium globosum / Heat shock proteins (HSPs) / Gene cloning and Expression

References

[1] Daniel C., John H.A. Evidence for the role of antibiosis in the antagonism of Chaetomium globosum to the apple scab pathogen, Venturia inaequalis [J]. Canadian Journal of Biology, 1984, 62: 1814-1818.

[2] Femando P., Heikkila J.J. Functional characterization of Xenopus small heat shock protein, Hsp30C: the carboxyl en is required for stability and chaperone activity [J]. Cell Stress & Chaperones, 2000, 5(2): 148-159.

[3] Fu X., Chang Z. Temperature-dependent subunit exchange and chaperone-like activities of Hsp16.3, a small heat shock protein from Mycobacterium tuberculosis [J]. Biochem Biophys Res Commun., 2004, 316(2): 291-299.

[4] Heikkila J.J. Expression and function of small heat shock protein genes during Xenopus development [J]. Semin Cell Dev Biol, 2003, 14(5): 259-266.

[5] Gu R., Fonseca S., Puskas L.G. et al. Transcript identification and profiling during salt stress and recovery of Populus euphratica [J]. Tree Physiol, 2004, 24(3): 265-276.

[6] Kanokmedhakul S., Kanokmedhakul K., Phonkerd N. et al. Antimycobacterial anthraquinone-chromanone compound and diketopiperazine alkaloid from the fungus Chaetomium globosum KMITL-N0802 [J]. Planta Med, 2002, 68(9): 834-836.

[7] Klein G., Laskowska E., Taylor A., Lipinska B. IbpA/B small heat-shock protein of marine bacterium Vibrio harveyi binds to proteins aggregated in a cell during heat shock [J]. Mar Biotechnol. (NY), 2001, 3(4): 346-354.

[8] Lentze N., Studer S., Narberhaus F. Structural and functional defects caused by point mutations in the alpha-crystallin domain of a bacterial alpha-heat shock protein [J]. J Mol Biol, 2003, 328(4): 927-937.

[9] Plesofsky N., Brambl R. Analysis of interactions between domains of a small heat shock protein, Hsp30 of Neurospora crassa [J]. Cell Stress Chaperones, 2002, 7(4): 374-386.

[10] Reissinger A., Winter S., Steckelbroeck S. et al. Infection of barley roots by Chaetomium globosum: evidence for a protective role of the exodermis [J]. Mycol Res, 2003, 107(Pt9): 1094-1102.

[11] Riou C., Nicaud J.M., Barre P. et al. Stationary-phase gene expression in Saccharomyces cerevisiae during wine fermentation [J]. Yeast, 1997, 13(10): 903-915.

[12] Sambrook J., Frisch E.F., Maniatis T. Molecular Cloning: A Laboratory Manual [M], 2002 Beijing: Science Press 485

[13] Sugiyama Y., Suzuki A., Kishikawa M. et al. Muscle develops a specific form of small heat shock protein complex composed of MKBP/HSPB2 and HSPB3 during myogenic differentiation [J]. J Bio-Chem., 2000, 275(2): 1095-1104.

[14] Usui K., Hatipoglu O.F., Ishii N. et al. Role of the N-terminal region of the crenarchaeal sHsp, StHsp14.0, in thermal-induced disassembly of the complex and molecular chaperone activity [J]. Biochem Biophys Res Commun, 2004, 315(1): 113-118.

[15] Wang Y., Bo H., Yang C. A method for rapid isolation of total RNA from Tamarix and Populus euphratica Oliv [J]. Journal of Northeast Forestry University, 2003, 31(5): 99-100.