Physical map of the Bacillus subtilis 166 genome: evidence for the inversion of an approximately 1900 kb continuous DNA segment, the translocation of an approximately 100 kb segment and the duplication of a 5 kb segment

Abstract

Physical map of theAn I-Ceul-Notl-Sfil endonuclease map of the Bacillus subtilis 166 genome was constructed. It was almost identical to that of B. subtilis 168 except for the inversion of an approximately 1900 kb DNA segment, the translocation of an approximately 100 kb segment and the duplication of a 5 kb segment. Continuity of the inverted segment was investigated by direct measurement of the distances between the two genomic loci where I-Scel recognition sites were created in the 168 and the 166 genomes. Size difference of the I-Scel fragments between the two strains fully demonstrated the inversion of an approximately 1900 kb long 'continuous' DNA segment and the 'location' of the two inversion junctions in the genome. The 100 kb DNA segment including the lysogenic SPp prophage was translocated close to one of the inversion junctions and was probably associated with the duplication of a 5 kb segment. These rearrangements are consistent with those indicated by genetic analyses. 1,

Construction of a combined physical and genetic map of the chromosome of Lactobacillus acidophilus ATCC 4356 and characterization of the rRNA operons

“…1). Chromosomal clustering of the rrn loci has not been found in L. plantarum (Kleerebezem et al, 2003), but it has been described for L. gasseri (Abs EL-Osta et al, 2002), L. sakei (Dudez et al, 2002) and Lactococcus lactis (Tulloch et al, 1991;Le Bourgeois et al, 1992) and for prokaryotes other then the lactic acid bacteria, such as E. coli, Bacillus subtilis, Haemophilus influenzae, Pasteurella multocida and Streptococcus thermophilus (Lee et al, 1989;Bentley et al, 1991;Roussel et al, 1994;Itaya, 1997;Hunt et al, 1998). In these bacteria, the rrn loci are often located within one-third to one-half of the chromosome.…”

Bacterial sources and sinks of isoprene, a reactive atmospheric hydrocarbon

“…This and other recent evidence (Wagner, 1998) suggest that strain 168 contains additional mutations that suppress the second and third phases of isoprene formation. Possibly, the chromosome of strain 168 underwent signi®cant chromosomal rearrangements during its isolation from the Marburg strain (Itaya, 1997). Further work should reveal the nature of these other mutations in strain 168 and how they suppress phases 2 and 3 of isoprene formation.…”

Sequential Insertion of Multiple I-SceI Recognition Sites at Designed Loci of theBacillus subtilis168 Genome

“…Instead, two commercially available endonucleases, I-SceI (5 0 -TAG-GGATAA/CAGGGTAAT-3 0 ) and I-PpoI (5 0 -ATGAC-TCTCTTAA/GGTAGCCAAA-3 0 ) were chosen due to their absence in the host genome and most of the guest genomes, although I-PpoI has been utilized to verify guest genome structures in the BGM vectors, 1,2) I-SceI remains of limited use. 8,9) Hence multiple I-SceI sites up to six were created in the host genome in this study. Six Sse8387I sites in the B. subtilis genome were selected ( Fig.…”

“…Two antibiotic resistance gene cassettes, neomycin (neo with I-SceI) and spectinomycin (spc with I-SceI), were constructed to create sequences at the two designed genome loci. 8,9) But this method apparently required numbers of antibiotic resistance genes equal to those of the I-SceI site to be created. Seven other antibiotic resistance gene cassettes are currently available for B. subtilis selection: chloramphenicol (cat), tetracycline (tet), erythromycin (erm), blasticidin S (bsr), kanamycin (kam), phleomyicn (phl), and hygromycin (hyg).…”