Gene cloning, expression, and characterization of recombinant aerolysin from Aeromonas hydrophila.

Abstract

Aerolysin is a significant virulent toxin protein secreted by Aeromonas hydrophila; it produces deep wound infections and hemorrhagic septicemia. The complete aerolysin gene (1,482 bp) was amplified from A. hydrophila. Furthermore, it was cloned and expressed into Escherichia coli BL21(DE3) codon plus RP cells using 0.5 mM IPTG for induction. The protein size was 54 kDa as estimated by SDS-PAGE, and it was purified by Ni-NTA affinity chromatography. Anti-His antibodies were used to characterize the expressed aerolysin by Western blotting and showed hemolytic activity with fish red blood cells. Aerolysin may be used as immunoassays for earlier control of A. hydrophila and is also compatible for vaccination.

• Authors: Vijai Singh, Pallavi Somvanshi, Gaurav Rathore, D Kapoor, B N Mishra
• Journal: Applied biochemistry and biotechnology (Appl Biochem Biotechnol) Vol. 160 Issue 7 Pg. 1985-91 (Apr 2010) ISSN: 1559-0291 [Electronic] United States
• PMID: 19763901 (Publication Type: Journal Article)

Chemical References

• Bacterial Toxins
• Pore Forming Cytotoxic Proteins
• Recombinant Proteins
• aerolysin

Topics

• Aeromonas hydrophila (genetics)
• Animals
• Bacterial Toxins (genetics, isolation & purification, metabolism)
• Blotting, Western
• Cloning, Molecular
• Fishes (blood, microbiology)
• Gene Expression
• Hemolysis
• Polymerase Chain Reaction
• Pore Forming Cytotoxic Proteins (genetics, isolation & purification, metabolism)
• Recombinant Proteins (genetics, isolation & purification)