EXPRESSION OF CONSERVED DOMAIN OF P1 PROTEIN OF M. PNEUMONIAE IN E.COLI

Background and Aim:Mycoplasma pneumoniae is a common pathogen that causes upper and lower respiratory tract infections in people of all ages, responsible for up to 40 % of community-acquired pneumonias. It also causes a wide array of extra pulmonary infections and autoimmune phenomena. P1 protein is a surface protein on the M. pneumoniae which are functional in receptor recognition, as the probable adhesion protein.Methods:Full length of P1 protein were analysed by bioinformatics tools. All of 93 P1 protein sequences available in NCBI data base were compared by multiple alignment tools. Consensus and hypervariable regions of P1 proteins were identified. Bioinformatics analysis identified four conserved and three hypervariable regions in 1628aa of P1 protein. All of important B-cell and T-cell epitopes were located only in two conserved b and d regions. So we chosed a highly conserved region residues 580 to 840 for experimental. For suitable expression in E. coli, all Codons were optimized in complete coding target peptide with 360 aa. Nucleotide sequence was cloned to pET32a+ vector. Expression of conserved domain of p1 protein of Mycoplasma pneumoniae was optimized in E. coli.Results:Results showed that the recombinant partial p1 protein fusion with TRX tag sequence was expressed in Bl21 strain of E. coli successfully.Conclusion:This peptide might be useful to developing a elisa diagnostic test.