Sequencing and characterization of plasmid pUIBI-1 from Bacillus thuringiensis serovar entomocidus LBIT-113.
Plasmid pUIBI-1 from Bacillus thuringiensis svr. entomocidus was sequenced and its replication mechanism analyzed. Sequence analysis revealed that pUIBI-1 contains 4671 bp and a 32% GC content. Plasmid pUIBI-1 also includes at least seven putative open reading frames (ORFs) encoding for proteins ranging from 5 to 50 kDa. ORF-1 encodes for a putative 16-kDa Rep protein, which lacks homology with proteins of similar function. ORF2 encodes for a protein of 50 kDa and shows homology with Mob proteins of plasmids pLUB1000 from Lactobacillus hilgardii (32.2%) and pGI2 from B. thuringiensis (33.7%). Detection of single-stranded DNA (ssDNA) intermediates indicated that pUIBI-1 replicates by the rolling-circle replication mechanism, as demonstrated by S1 treatment and Southern hybridization under non-denaturing conditions.
López-Meza
Joel E
JE
Centro Multidisciplinario de Estudios en Biotecnología-FMVZ, Univ. Michoacana de San Nicolás de Hidalgo, Apdo. Postal 53, Administración Chapultepec, 58262 Morelia, Michoacán, México.
Barboza-Corona
J Eleazar
JE
Del Rincón-Castro
M Cristina
MC
Ibarra
Jorge E
JE
D001413:Bacillus thuringiensis / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics; D001482:Base Composition; D026501:Blotting, Southwestern / Q000379:methods; D004261:DNA Replication; D004277:DNA, Single-Stranded / Q000032:analysis; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010957:Plasmids / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000378:metabolism
D001426:Bacterial Proteins; D004277:DNA, Single-Stranded; C083180:Mob protein, Bacteria; D015719:Single-Strand Specific DNA and RNA Endonucleases