Tn5-mediated integration of the delta-endotoxin gene from Bacillus thuringiensis into the chromosome of root-colonizing pseudomonads
| Autor: | F. J. Perlak, M. G. Obukowicz, S L Bolten, K Kusano-Kretzmer, E J Mayer, L S Watrud |
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| Rok vydání: | 1986 |
| Předmět: |
Bacterial Toxins
Bacillus thuringiensis DNA Recombinant Transposases Pseudomonas fluorescens Microbiology Hemolysin Proteins fluids and secretions Bacterial Proteins Insertion Molecular Biology Gene Transposase Genetics Bacillus thuringiensis Toxins biology Genetic transfer Nucleic Acid Hybridization Chromosomes Bacterial biology.organism_classification Nucleotidyltransferases Molecular biology Endotoxins Genes Bacterial Mutation Horizontal gene transfer DNA Transposable Elements bacteria Research Article Delta endotoxin |
| Zdroj: | Journal of Bacteriology. 168:982-989 |
| ISSN: | 1098-5530 0021-9193 |
| DOI: | 10.1128/jb.168.2.982-989.1986 |
| Popis: | Gene replacement mediated by Tn5 sequences was used to integrate the Bacillus thuringiensis subsp. kurstaki HD-1 delta-endotoxin gene (tox) into the chromosome of two corn root-colonizing strains of Pseudomonas fluorescens. A Tn5 transposase deletion element containing the tox gene (delta Tn5-tox) was substituted for a Tn5 element previously present in the P. fluorescens chromosome. Two classes of delta Tn5-tox elements were made. The first class encodes kanamycin resistance in addition to the Tox protein, whereas the second class encodes only the Tox protein. Both classes of delta Tn5-tox elements can no longer transpose, owing to a 324-base-pair deletion in the transposase gene of IS50R, minimizing the potential for horizontal gene transfer of the tox gene to other bacterial species. A frameshift mutation in the transposase gene of IS50L was also constructed to eliminate the possibility of suppression or of a spontaneous reversion at the ochre termination codon that would create an active transposase. Expression of the Tox protein in P. fluorescens strains 112-12 and Ps3732-3-7 was demonstrated by an immunological assay (Western blot) and toxicity against larvae of the tobacco hornworm (Manduca sexta). |
| Databáze: | OpenAIRE |
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