- Volume 140, Issue 9, 1994
Volume 140, Issue 9, 1994
- Genome Analysis
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Determination of genome size and a preliminary physical map of an extreme alkaliphile, Micrococcus sp. Y-1, by pulsed-field gel electrophoresis
More LessLarge restriction fragments of genomic DNA from Micrococcus sp. Y-1 were separated by pulsed-field gel electrophoresis (PFGE). Since Micrococcus sp. Y-1 has a G + C content of approximately 70%, restriction fragments were obtained by digesting chromosomal DNA with endonucleases which recognize A+T-rich sequences. Five enzymes, Sspl, Spel, Xbal, Hpal and EcoRI, were used for generation of distinctly separated fragments in the size range 100-500 kb. No site for DraI was detected. In contrast, sites for 8-base-recognizing enzymes, but not for NotI and Sfil, were frequent. The genome size of Micrococcus sp. Y-1 was determined from restriction fragments separated by PFGE, and was estimated to be approximately 4061 kb. Partial digestion experiments revealed the order of the six Sspl fragments on the chromosome.
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Cloning and nucleotide sequencing of a 15 kb region of the Bacillus subtilis genome containing the iol operon
More LessWithin the framework of an international project on the sequencing of the whole Bacillus subtilis genome, a 15 kb chromosome segment, which contains the iol operon involved in inositol utilization, has been cloned and sequenced. This region (14974 bp) contains 12 complete open reading frames (ORFs; genes) and two partial ones; the seventh gene (E83G) is the idh gene encoding inositol dehydrogenase. All the genes identified are transcribed in the same direction as that of the movement of the replication fork. A homology search for their products deduced from the 12 complete ORFs revealed that eight of them exhibit significant homology to known proteins such as fructokinase, acetolactate synthase, fructose-1,6-bisphosphate aldolase (B. subtilis), and PhoB and FtsE proteins (Escherichia coli). It also implied that two genes (B65B and B65E) might encode a set of two-component regulatory proteins and that the B65F gene might encode a protein belonging to the ATP-binding cassette (ABC) family. Based on the features of the nucleotide sequence determined and the results of the homology search, the primary structure of the iol operon is predicted.
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Two identical copies of IS1246, a 1275 base pair sequence related to other bacterial insertion sequences, enclose the xyl genes on TOL plasmid pWWO
More LessTwo identical direct repeats of a 1275 bp sequence, designated IS1246, encompass the xyl genes, which determine the catabolism of toluene, m- and p-xylenes to central metabolites, on the TOL catabolic plasmid pWWO. IS1246 has a terminal inverted repeat of 12 bp (5‘GGGCACCTCGAA3’) and contains a major open reading frame of 280 codons. This ORF shows significant homology with ORFs encoded by a number of bacterial insertion sequences from Bacteroides, Neisseria and Escherichia coli.
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