Cloning, expression and characterization of aerolysin from Aeromonas hydrophila in Escherichia coli.

Abstract

Aerolysin is a toxin (protein in nature) secreted by the strains of Aeromonas spp. and plays an important role in the virulence of Aeromonas strains. It has also found several applications such as for detection of glycosylphosphatidylinositol (GPI)-anchored proteins etc. A. hydrophila is a ubiquitous Gram-negative bacterium which causes frequent harm to the aquaculture. To obtain a significant amount of recombinant aerolysin in the active form, in this study, we expressed the aerolysin in E. coli under the control of T7 RNase promoter. The coding region (AerA-W) of the aerA gene of A. hydrophila XS91-4-1, excluding partial coding region of the signal peptide was cloned into the vector pET32a and then transformed into E. coli b121. After optimizing the expression conditions, the recombinant protein AerA-W was expressed in a soluble form and purified using His. Bind resin affinity chromatography. Recombinant aerolysin showed hemolytic activity in the agar diffusive hemolysis test. Western blot analysis demonstrated good antigenicity of the recombinant protein.

• Authors: Daling Zhu, Aihua Li, Jianguo Wang, Ming Li, Taozhen Cai
• Journal: Indian journal of biochemistry & biophysics (Indian J Biochem Biophys) Vol. 44 Issue 4 Pg. 204-8 (Aug 2007) ISSN: 0301-1208 [Print] India
• PMID: 17970277 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Bacterial Toxins
• Pore Forming Cytotoxic Proteins
• Protein Sorting Signals
• Recombinant Proteins
• aerolysin

Topics

• Aeromonas hydrophila (metabolism)
• Bacterial Toxins (immunology, isolation & purification)
• Chromatography, Affinity
• Cloning, Molecular (methods)
• Escherichia coli (genetics, metabolism)
• Pore Forming Cytotoxic Proteins (immunology, isolation & purification)
• Protein Sorting Signals (genetics, physiology)
• Recombinant Proteins (isolation & purification, metabolism)