I-CeuI recognition sites in the rrn operons of the Bacillus subtilis 168 chromosome: inherent landmarks for genome analysis.
The Bacillus subtilis 168 circular chromosome yielded ten fragments on I-CeuI endonuclease digestion. I-CeuI recognizes a 26 bp sequence that is located within the gene encoding the 23S subunit of the rRNA in Chlamydomonas eugametos, Escherichia coli and Salmonella typhimurium. The precise locations of the I-CeuI sites of the B. subtilis chromosome were determined on a NotI-SfiI physical map by (i) double digestion analyses with I-CeuI and SfiI, (ii) comparison of mutant strains lacking a specific rrn operon, (iii) using an I-CeuI linking clone and (iv) analysis of nucleotide sequence data of some rrn operons. In conclusion, all the I-CeuI sites were located within the B. subtilis rrn operons and the I-CeuI sites were conserved in all the B. subtilis 168 derivatives tested. Thus, variations in size of the I-CeuI fragments must be due to genome alterations. A B. subtilis 168 strain was investigated with I-CeuI. We demonstrated that the aberrant structure was the outcome of the inversion of an approximately 1700 kb DNA segment.
Toda
Tsutomu
T
School of Science and Engineering, Saitama University, Urawa-shi, Saitama 338, Japan.
Itaya
Mitsuhiro
M
Mitsubishi Kasei Institute of Life Sciences, 11 Minamiooya, Machida-shi, Tokyo 194, Japan.
D001412:Bacillus subtilis / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D007446:Chromosome Inversion; D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial; D004269:DNA, Bacterial / Q000032:analysis; D015252:Deoxyribonucleases, Type II Site-Specific; D016521:Electrophoresis, Gel, Pulsed-Field; D004706:Endodeoxyribonucleases; D015698:Genomic Library; D008969:Molecular Sequence Data; D009154:Mutation; D012338:RNA, Ribosomal, 23S / Q000235:genetics*; D012407:rRNA Operon / Q000235:genetics*
D004269:DNA, Bacterial; D012338:RNA, Ribosomal, 23S; D004706:Endodeoxyribonucleases; C071132:endodeoxyribonuclease I-CeuI; C041749:endodeoxyribonuclease SfiI; D015252:Deoxyribonucleases, Type II Site-Specific; C038565:GCGGCCGC-specific type II deoxyribonucleases