Production and Purification of the Peptidoglycan-associated Lipoprotein (Pal) of Legionella pneumophila

R.A. Rajabi¹, A. MohabatiMobarez¹*, Ali HatefSalmanian², N. Khoramabadi¹ and H. Aghababa¹

¹TarbiatModares University, Faculty of Medical Sciences, Department of Bacteriology, Tehran, Iran. ²Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology(NIGEB), Shahrak-e-Pajoohesh, 15thKm, Tehran-Karaj Highway, Tehran, Iran.

DOI : http://dx.doi.org/http://dx.doi.org/10.13005/bbra/1030

ABSTRACT:

Today, Legionella pnuemophila is identified as usual cause of both community and nosocomial pneumonia. The PAL (peptidoglycan-associated lipoprotein) of Legionella is the most prominent surface antigen, widely conserved,species-common, immune-dominant and potentialurinary diagnostic antigen.PAL potentially elicits immune responses via the interaction with TLR2-harboring cells.Isolation, cloning and expression of PAL were aimed in this research. Pal geneof Legionella pnumophila serogroup 1, strain Paris, was amplified by PCR method,sub cloned into pET28a [+) as expression vector and sequenced in both directions. Recombinant protein after over-expression in E.coli strains BL21 (DE3) and purified by an affinity purification process with Ni+2 –nitrilotriacetic acid (NTA)-agarose. Sequencing results and western blot analysis with anti His tag,rabbit polyclonal anti-L.pneumophila antiserum and L.pneumophila-infected patient serum antibody confirmed the successful expression of recombinant PAL in the heterologous host E.coli.

KEYWORDS: pal; Legionella pnuemophila; Recombinant PAL.

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