FIG. 2.

Identification of the P48 gene in the M. agalactiae genome. (A) Typical results obtained by Southern analysis. Genomic DNA (5 μg/well) was digested with BglII and EcoRI (lane 1), EcoRI (lane 2), XbaI (lane 3), PstI (lane 4), HindIII (lane 5), and PstI and HindIII (lane 6). After electrophoresis and capillary transfer to a positively charged nylon membrane, digested DNA was hybridized with a digoxigenin-labeled oligoprobe derived from the N-terminal microsequencing of P48. Two overlapping fragments, namely the 4-kb HindIII and 6.6-kb PstI fragments, were gel purified, cloned in pUC18, and partially sequenced. (B) Physical map and partial characterization of coding regions corresponding to the 4-kb HindIII and 6.6-kb PstI overlapping fragments. The P48 gene is a malp (7) homolog and encodes a 22-residue leader peptide (white box), followed by the mature P48 lipoprotein (right dashed box). Another ORF (left dashed box) was found in opposite orientation and encodes a putative 59-kDa homolog of the P63 ABC transporter of M. fermentans. H, P, and E indicate HindIII, PstI, and EcoRI restriction sites, respectively.