Analysis of the Bacillus subtilis genome: cloning and nucleotide sequence of a 62 kb region between 275 degrees (rrnB) and 284 degrees (pai).
In the framework of the international project aimed at the sequencing of the Bacillus subtilis genome, five DNA fragments in the region between rrnB (275 degrees) and pai (284 degrees) were cloned by inverse and combinatorial long-range PCR and their nucleotide sequences were determined and analysed. Together these sequences constituted a contig of 62229 bp. On the basis of the position of Not1 and Stil restriction sites, the orientation and order of known genetic markers was determined to be pai (284 degrees)-degQ comQ comP comAA comAB-pbpD-kapB kinB patB-mcpB tipA mcpA tipB-rrnB (275 degrees). Fifty-four ORFs were detected. Thirteen of these coincided with known B. subtilis genes, and 41 new ORFs were found. Of the predicted new gene products, 12 showed no significant similarity to other known proteins, whereas ten showed strong similarity to proteins of other organisms with unknown function. Nineteen predicted proteins showed strong similarity to known proteins of other organisms, for instance a Na+/H+ antiporter system of Bacillus alcalophilus, a sugar transport system found in Mycoplasma genitalium, NADH-dependent butanol dehydrogenase of Clostridium acetobutylicum, glucose-6-phosphate isomerase A of B, subtilis, exo-1,4-alpha-glucosidase activity of Bacillus stearothermophilus and L-rhamnose isomerase of Escherichia coli.
Oudega
Bauke
B
Department of Molecular Microbiology, Institute of Molecular Biological Sciences, BioCentrum Amsterdam, Faculty of Biology, Vrije Universiteit, De Boelelaan 1087, 1081 HV Amsterdam, The Netherlands.
Koningstein
Gregory
G
Department of Molecular Microbiology, Institute of Molecular Biological Sciences, BioCentrum Amsterdam, Faculty of Biology, Vrije Universiteit, De Boelelaan 1087, 1081 HV Amsterdam, The Netherlands.
Rodrigues
Luísa
L
Department of Molecular Microbiology, Institute of Molecular Biological Sciences, BioCentrum Amsterdam, Faculty of Biology, Vrije Universiteit, De Boelelaan 1087, 1081 HV Amsterdam, The Netherlands.
de Sales Ramon
Maria
M
Department of Molecular Microbiology, Institute of Molecular Biological Sciences, BioCentrum Amsterdam, Faculty of Biology, Vrije Universiteit, De Boelelaan 1087, 1081 HV Amsterdam, The Netherlands.
Hilbert
Helmut
H
Qiagen GmbH, Max-Volmer Strasse 4, D-40724 Hilden, Germany.
Düsterhöft
Andreas
A
Qiagen GmbH, Max-Volmer Strasse 4, D-40724 Hilden, Germany.
Pohl
Thomas M
TM
Gesellschaft für Analyse-Technik und Consulting GmbH, Fritz-Arnold Strasse 23, D-78467 Konstanz, Germany.
Weitzenegger
Thomas
T
Gesellschaft für Analyse-Technik und Consulting GmbH, Fritz-Arnold Strasse 23, D-78467 Konstanz, Germany.
D001412:Bacillus subtilis / Q000235:genetics*; D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial / Q000235:genetics; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D016680:Genome, Bacterial; D007391:International Cooperation; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D012984:Software