Bacillus subtilis is the best-characterized member of the Gram-positive bacteria. Its genome of 4,214,810 base pairs comprises 4,100 protein-coding genes. Of these protein-coding genes, 53% are represented once, while a quarter of the genome corresponds to several gene families that have been greatly expanded by gene duplication, the largest family containing 77 putative ATP-binding transport proteins. In addition, a large proportion of the genetic capacity is devoted to the utilization of a variety of carbon sources, including many plant-derived molecules. The identification of five signal peptidase genes, as well as several genes for components of the secretion apparatus, is important given the capacity of Bacillus strains to secrete large amounts of industrially important enzymes. Many of the genes are involved in the synthesis of secondary metabolites, including antibiotics, that are more typically associated with Streptomyces species. The genome contains at least ten prophages or remnants of prophages, indicating that bacteriophage infection has played an important evolutionary role in horizontal gene transfer, in particular in the propagation of bacterial pathogenesis.

The 49630 bp sjpoOil-rmH region of the Bacillus subtilis genome has been fully sequenced. The sequence contains one partial and 62 complete ORFs, one partial and three complete rRNA genes and a cluster of six tRNA genes. The direction of the transcription and translation of 61 ORFs is the same as that of the movement of the replication fork. A homology search of 40 ORFs in newly determined sequence revealed that 27 of them had significant similarity to known proteins such as elongation factor G, elongation factor Tu, pseudouridine synthase I and ribosomal proteins. Two adjacent genes, ybaD and yh€, appeared to encode proteins belonging to the ATP-binding cassette (ABC) family.Keywords : Bacilhs stlbtiljs, genome sequencing, ribosomal protein gene cluster INTRODUCTIONWithin the framework of the international project for the sequencing of the entire Bacdlltls stlbtdh genome, we are responsible for sequencing a chromosomal region of about 150 kb between SpoOH(10") and pNEXT9 (24"). In this study, we determined the sequence of about 30 kb in the region between 10" and 15". We combined the results with known sequences and describe here a continuous 49630 bp sequence from spool3 (10') to rrtaH (15'). METHODSBacterial strains, phage and plasmids. B. subtilis 168 (trpC2), from our laboratory stock, was used as a template for PCR.Escherichia coli DH5a (stlpE44 lacU169 [480 lacZAM151 hsdR 17 recAl endAl gyrA96 tbi-I relAI) was used as the host for cloning with plasmid pTZ18R (Toyobo). The 1 phage K4F811 derived from EMBL 3a was supplied by F. Morohoshi (National Cancer Center, Tokyo, Japan).Sequence determination. All PCR products to be sequenced were self-ligated and fragmented by sonic oscillation, then 400-700 bp fragments were purified by means of agarose gel electrophoresis. These fragments were cloned into a vector (pTZ18R) and randomly overlapping libraries for shotgun sequencing were prepared. Inserts of pTZl8R were amplified by PCR using a forward (5' CGTCAAGGCGATTAAGTTG-GGTAA 3') and a reverse (5' TGCTTCCGGCTCGTATGT-TGTGTG 3') sequencing primer based on the vector sequence. The amplified PCR products were purified by precipitation with polyethylene glycol and used as templates for sequencing. Sequencing reactions were performed on both strands using the CATALYST Molecular Biology Labstation -Model 800 and Taq Dye Primer Cycle Sequencing Kit (Applied Biosystems), and were then analysed with a Model 373A DNA Sequencer (Applied Biosystems).Data handling and computer analysis. DNA sequences were compiled using the GENETYX-MAC ATSQ (Software Development) and GENEWORKS programs (IntelliGenetics), We used sequence outputs only from the regions where every base was well separated (about 400 bp in one run). The compiled sequence was further searched for possible ORFs using GENETYX-MAC and GENEWORKS. A homology search of the identified ORFs was performed with the FASTA program using GENETYX-CD (Software Development) and a GenomeNet FASTA server in the Human Genome Center, Institute of Medical Science, University of Tokyo,...

No abstract

A 31 141 bp continuous nucleotide sequence in the region from trnl to pNEXT52 in the Bacillus subtilis 168 genome was determined. In the region, there were 22 ORFs, two complete rRNA operons, and five tRNA genes. It was deduced that the function of one of the ORFs was similar to that of a sigma factor belonging to the ECF (extra-cytoplasmic functions) subfamily. The gene cluster feuA, B, C reported previously for other strains of B. subtilis was also found in strain 168 and located in this region.

Three patients with hepatic cirrhosis who had undergone gastric operations following endoscopic sclerotherapy were retrospectively reviewed. One patient had undergone total gastrectomy for gastric cancer and two had Hassab and Tanner operations for gastric and esophageal varices. All patients were recovering with no complications related to the operations and were free of disease for 2 years, postoperatively.When esophageal varices are found in combination with an upper gastric cancer that requires total gastrectomy, endoscopic sclerotherapy for esophageal varices is contraindicated because severe esophageal injuries may be induced by the sclerosing agent. The Hassab and Tanner method is useful for esophageal and gastric varices after sclerotherapy. A repeat endoscopic sclerotherapy should be performed after this operation.