Initiation of mini-ftsZ RNA transcripts in B. subtilis. The B. mycoides mini-ftsZ DNA construct was cloned into pJPR1 and inserted at the AmyE site of B. subtilis 168 (see methods). Transcripts of the construct were detected in total B. subtilis RNA by primer extension from the labeled primer Amy5 (Table 1) specific to the amyE 5’ region located 245 nt downstream of the inserted construct. A) Autoradiogram of PE. Lanes1 and 2: transcripts originating from the Pxyl promoter, induced by 5% xylose for 18 and 3 hours. Lane 3: the faint transcripts of the ftsZ minigene present in the non-induced B. subtilis recombinant strain are indicated by asterisks and map at −140 and −10 from the first nucleotide of the minigene ftsZ ORF as in B. mycoides. These bands are not present in the control B. subtilis strain (lane 4). B) schematic view of the construct in pJPR1. C) Schematic representation of the cDNAs indicated by asterisks in A. The red circle marks the position of the terminator structure 3’ to the B. mycoides ftsZ ORF. M = MW marker DNA. GATC = M13MP18 sequence ladder.
Santini et al. BMC Microbiology 2013 13:27 doi:10.1186/1471-2180-13-27