Genetics and Biotechnology of Bacilli by A. T. Ganesan

By A. T. Ganesan

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Mutations which change the length of the spacer of E. , 1985). In a survey of B. subtilis 43 Ε σ promoters, Moran et al. (1982) observed strong conservation of the sequence 5'-RTRTG-3', usually found at positions -18 to -14 upstream from the site of transcription initiation; this sequence will be referred to here as the -16 region. The T-TG part of this sequence is a weakly conserved element in E. coli promoters (Hawley and McClure, 1983). The tms promoter, a vegetative B. , 1982), shows good adherence to consensus sequences at the -35 and -10 regions (5/6 matches at each) and has the preferred 17 bp spacing between these sequences, but has only 3/5 adherence to the RTRTG sequence in the -16 region (Figure 1).

We next examined the level of mRNA for aprE by SI mapping analysis. An SI-protected DNA fragment of an expected size (520 bases) was much more abundant in the RNA sample obtained from the cells carrying prtR on pNC6 (Fig. 3c), as compared to the band in the RNA sample obtained from the cells carrying pNC6 alone (Fig. 3b). The result indicates that prtR increases the level of mRNA of the apr gene. The increased mRNA level could be caused by an enhanced rate of mRNA synthesis or increased mRNA stability.

J. Henner , M. A. , 460 Point San Bruno Blvd, South San Francisco, CA 94080; ^Research Institute of Scripps Clinic, 10666 North Torrey Pines Rd, La Jolla, CA 92037 1 I. INTRODUCTION The onset of sporulation in Bacillus subtilis triggers the expression of a set of genes, some of which are essential to the sporulation process and other that are only coincidental to it. The expression of the alkaline protease (aprE) gene, which belongs to the class of nonessential sporulation associated genes (Stahl and Ferrari 1984), represents a very exquisite tool to study how this differentiation process is controlled at the transcriptional and/or translational level.

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