Figure 2

SDS-PAGE and Western blot analysis of different fractions of B. halodurans recombinant strains transformed with plasmid pSECFliCFFuz. (A) SDS-PAGE (12%) analysis of 16 h crude cell-culture supernatants (20 μl) and intracellular fractions (20 μl) from the different B halodurans strains as indicated above each lane. (B) Western blot analysis of the same samples as in A using Anti-FLAG monoclonal antibodies. The letter S indicates cell-free culture supernatant and I intracellular fraction. (C) Western blot analysis with an antibody to chloramphenicol acetyltransferase as a test for cell lysis. The crude cell-free supernatants (S) are the same as in A and the intracellular fractions are from control strain BhFDL05S (pSECFliCFFuz), lane 1; BhFDL05S (pSEC194), lane 2 and negative control BhFDL05S without plasmid, lane 3. The arrow indicates the expected size for chloramphenicol acetyltransferase (35 kDa). The molecular weight markers in kDa are indicated on the left.