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Genomics |
Department of Biotechnology, Faculty of Engineering, Fukuyama University, 985 Sanzo, Higashimura-cho, Fukuyama 729-0292, Japan1
Author for correspondence: Y. Fujita. Tel: +81 849 36 2111. Fax: +81 849 36 2459. e-mail: yfujita{at}bt.fubt.fukuyama-u.ac.jp
Within the framework of the international project ‘The functional analysis of the Bacillus subtilis genome’ in Japan and Europe, the gene expression and transcription organization of the gntZ–ywaA region (160 kb) of the B. subtilis genome has been systematically analysed. First, all unanalysed genes comprising more than 80 amino acids (125 genes) in this region were inactivated through integration of plasmid pMUTIN. No essential gene was found which could not be inactivated. All the integrants grew normally in both nutrient sporulation medium and glucose minimal medium. But an integrant in the yxbG gene exhibited an oligosporogenic phenotype in the nutrient sporulation medium. The synthesis of ß-galactosidase was examined, as a reporter for expression of the inactivated genes, during growth and sporulation in the two media. The results indicated that 36% of the promoters were inactive when cells were grown in at least one of these two media. Furthermore, the transcription of the 119 genes in this region was analysed by Northern blotting, resulting in a transcription map. The results indicate that the gntZ–ywaA region contains at least 24 polycistronic operons, including several published ones. The operons newly found in this work are yxaAB, yxaGH, yxaJKL, yxbBA–yxnB–asnH–yxaM, yxbCD, yxcED, yxdJK, yxeFGH, yxeKLMNOPQ, yxeR–yxxB, hutPHUIGM, bglPH–yxiE, wapA–yxxG, yxiM–deaD, katB–yxiS, yxjCDEF, yxjJI and yxkF–mmsX.
Keywords: Bacillus subtilis, genome, transcription, gene inactivation, gene expression
Abbreviations: ß-Gal, ß-galactosidase; DSM, defined nutrient sporulation medium; MM, glucose minimal medium; SD, Shine–Dalgarno
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