Fig. 7.

Triton X-100 extraction of Lig proteins from L. kirschneri RM52. Leptospires were incubated for 2 h in MEM (ATCC) at 37°C. Bacteria were then washed once in 1× PBS with 5 mM MgCl2 and extracted with 1% Triton X-100. The insoluble protoplasmic cylinder (PC) was pelleted by centrifugation. The protoplasmic cylinder and the Triton-soluble (SOL) fractions were probed in a Western blot using LigA/B repeat (1:5000), LipL41 (1:10 000), and LipL31 (1:8000) antisera. LipL31 is a marker for the leptospiral inner membrane (Haake and Matsunaga, 2002).