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

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. // //