FIGURE 1.
Detection of YycG and YycF in membrane and cytosolic protein fractions. Cultures of TIGR4 carrying either pLS1RGFP or pPL101 plasmids were induced and then, membrane and cytosolic protein extracts were prepared. (A) the extracts were [32P]–phosphorylated in vitro, fractionated by SDS-PAGE, and protein bands detected by autoradiography. (B) the extracts were fractionated by SDS-PAGE followed by western blotting and detection using antibody developed against YycF (see details in Materials and Methods). Position of migration of the SeeBlue Plus2 (Invitrogen) pre-stained protein standard are indicated. Phosphorylase B, 148 kDa; bovine serum albumin, 98 kDa; glutamic dehydrogenase, 64 kDa; alcohol dehydrogenase, 50 kDa; carbonic anhydrase 36 kDa; myoglobin red 22 kDa and lysozyme 16 kDa.