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Molecular Detection and Expression of Azurin: An Anti-Cancer Protein from Local Isolates of Pseudomonas species

Aslam F, Farooq A, Qaiser H, Bajwa S, Saleem F and Naz S

Azurin was recognized as anticancer drug in 2000, it plays an important role in inducing apoptosis and cytotoxicity. Azurin may incorporate with cytosol and nuclear material after penetrating the tumor cells and stabilize p53 (a tumor suppresser protein) that facilitate to increase its level inside the cell. A 447 bp PCR amplified fragment of Azurin from Pseudomonas aeruginosa was cloned in pTG19-T cloning vector. The DNA sequence predicts a pre-protein with a signal peptide of 19 amino acids followed by 128-aminoacid mature azurin protein. azurin gene was cloned in pET22b vector. Recombinant rpET22b (+)-Azu vector was cloned in BL21-Codon Plus (DE3) cells. Optimized, high expression of azurin gene was obtained at 0.5 mM concentration of IPTG after 6 hours at 37°C. Sequencing shows 99.8% sequence homology with reported sequence of azurin. One silent mutation i.e., CCA to CCG is detected at 17th amino acid. However, resulting amino acid (proline) remains the same.