Fig 5.
In vivo chemical cross-linking of B. subtilis cells with various ykn and sdpC genotypes. (A and B) In vivo DSP cross-linking of HB6127 (yknWXYZ∷Kan) cells carrying plasmids producing the indicated combinations of Ykn proteins. Membrane fractions were separated by 10% SDS-PAGE followed by immunoblotting with anti-YknW (A) and anti-YknX (B) antibodies. Molecular masses of high-molecular-weight cross-linked complexes are indicated. (C) HB6186 (Pspac-hy-sdpABC sdpI∷mls yknWXYZ∷Kan) cells carrying pHTWXYZ were grown in different combinations of 1 mM IPTG and 0.5% xylose to induce the expression of sdp or ykn. Cells were treated with DSP, and membrane fractions were analyzed by immunoblotting with the indicated polyclonal antibodies.