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Original Article

Identification and Characterization of the omaA Gene Encoding the Major Outer Membrane Protein of Azospirillum brasilense

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Pages 225-237 | Received 17 Mar 2000, Published online: 11 Jul 2009
 

Abstract

The major outer membrane protein (MOMP) of Azospirillum brasilense was purified and degenerate oligo-nucleotides were constructed on the basis of partial internal amino acid sequences. PCR products were obtained using total DNA of A. brasilense as template. One of these, a 766-bp fragment, was DIG-labelled and used in Southern hybridization against A. brasilense DNA and a genomic library of A. brasilrnsc in Escherichia coli. A clone containing a 20-kb EcoRI insert in pLAFR3 was identified by PCR screening. From this insert, an EcoRI-SalI fragment of approximately 3.5-kb was subcloned in pUC19. The gene encoding the A. brasilense MOMP was sequenced and analyzed. The deduced amino acid sequence contains a putative signal peptide of 23 residues, followed by 367 amino acids of the mature protein with a molecular mass of 38,753 Da. The deduced amino acid sequence shows similarity to certain bacterial porins.

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