Expression and purification of lectin a from pseudomonas aeruginosa in E.coli

Author(s): B. Veeresh, G. Adarsh, T. Ramesh, M. Ramesh*

Abstract

Pseudomonas aeruginosa is a pathogenic micro organism which infects cystic fibrosis patients. The mode of infection is by attaching to the host cell using lectin proteins present in cell wall of Pseudomonas aeruginosa. The genomic DNA has been isolated from Pseudomonas aeruginosa. The Lec A gene coding for lectin protein has been isolated from genomic DNA of Pseudomonas aeruginosa using specific primers by specific gene amplification using PCR. The isolated Lec A gene is cloned by T/A cloning. The Lec A gene is inserted into expression vector pET-32a. The recombinant vector is transformed into E. coli BL-21. The protein is over expressed by adding inducer IPTG. The expressed protein is purified by using affinity chromatography and confirmed by SDS PAGE.

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