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J. Biochem, 2003, Vol. 133, No. 4 475-483
© 2003 Japanese Biochemical Society

MOLECULAR BIOLOGY

Transcriptional, Functional and Cytochemical Analyses of the veg Gene in Bacillus subtilis

Tatsuya Fukushima1, Shu Ishikawa1, Hiroki Yamamoto1, Naotake Ogasawara2 and Junichi Sekiguchi+,1

1 Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, 3-15-1 Tokida, Ueda, Nagano 386-8567; and 2 Graduate School of Information Science, Nara Institute of Science and Technology, Ikoma, Nara 630-0101

A Bacillus subtilis veg mutant exhibited a small reduction of absorbance, a large reduction of hexosamine release, and slow dipicolinic acid release from spores during germination with L-alanine as a germinant. But veg spores exhibited normal resistance to chloroform, 2-propanol, lysozyme, and heat. SDS–polyacrylamide gel electrophoresis of spore coat proteins revealed no difference in coat proteins between the wild type and the veg mutant. Northern and veg-lacZ fusion analyses indicated that the veg gene is transcribed in both the vegetative growth and sporulation phases, and primer extension analysis indicated an identical transcriptional start point in both phases. The upstream sequence suggests that veg is transcribed by E{sigma}A RNA polymerase. Veg-GFP fusion protein was detected for vegetative cells and spores, but the fluorescence of mother cells disappeared completely in the late sporulation phase. Decoated spores containing Veg-GFP exhibited a strong green fluorescence in the core, but much weaker fluorescence in the cortex.

+ To whom correspondence should be addressed. Tel: +81-268-21-5344, Fax: +81-268-21-5345, E-mail: jsekigu{at}giptc.shinshu-u.ac.jp


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