Sequencing and Characterization of Plasmid pUIBI-1 from Bacillus thuringiensis Serovar entomocidus LBIT-113 |
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| Authors: | Joel E. López-Meza J. Eleazar Barboza-Corona M. Cristina Del Rincón-Castro Jorge E. Ibarra |
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| Affiliation: | (1) Centro Multidisciplinario de Estudios en Biotecnología-FMVZ, Univ. Michoacana de San Nicolás de Hidalgo, Apdo. Postal 53, Administración Chapultepec, 58262 Morelia, Michoacán, Mexico;(2) Instituto de Ciencias Agricolas, Universidad de Guanajuato, Apdo. Postal 311, 36500 Irapuato, Gto., Mexico;(3) Departamento de Biotecnología y Bioquímica, Centro de Investigación y de Estudios Avanzados del I.P.N., Apdo. Postal 629, 36500 Irapuato, Gto., Mexico |
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| Abstract: | 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. |
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