In vivo random mutagenesis of streptomycetes using mariner-based transposon Himar1
| العنوان: | In vivo random mutagenesis of streptomycetes using mariner-based transposon Himar1 |
|---|---|
| المؤلفون: | Stephen Weber, Bohdan Bilyk, Maksym Myronovskyi, Oksana Bilyk, Andriy Luzhetskyy, Lutz Petzke |
| المصدر: | Applied Microbiology and Biotechnology. 97:351-359 |
| بيانات النشر: | Springer Science and Business Media LLC, 2012. |
| سنة النشر: | 2012 |
| مصطلحات موضوعية: | Genetics, Microbial, Transposable element, Genetics, biology, Streptomyces coelicolor, General Medicine, biology.organism_classification, Sleeping Beauty transposon system, Applied Microbiology and Biotechnology, Streptomyces, Genomic Instability, Anti-Bacterial Agents, Mutagenesis, Insertional, Mutation Rate, Drug Resistance, Bacterial, DNA Transposable Elements, Recombinase, bacteria, Transposon mutagenesis, Selection, Genetic, Transposase, Streptomyces albus, Biotechnology |
| الوصف: | We report here the in vivo expression of the synthetic transposase gene himar1(a) in Streptomyces coelicolor M145 and Streptomyces albus. Using the synthetic himar1(a) gene adapted for Streptomyces codon usage, we showed random insertion of the transposon into the streptomycetes genome. The insertion frequency for the Himar1-derived minitransposons is nearly 100 % of transformed Streptomyces cells, and insertions are stably inherited in the absence of an antibiotic selection. The minitransposons contain different antibiotic resistance selection markers (apramycin, hygromycin, and spectinomycin), site-specific recombinase target sites (rox and/or loxP), I-SceI meganuclease target sites, and an R6Kγ origin of replication for transposon rescue. We identified transposon insertion loci by random sequencing of more than 100 rescue plasmids. The majority of insertions were mapped to putative open-reading frames on the S. coelicolor M145 and S. albus chromosomes. These insertions included several new regulatory genes affecting S. coelicolor M145 growth and actinorhodin biosynthesis. |
| تدمد: | 1432-0614 0175-7598 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::7a39bef88844a038f316250f409ffc60 https://doi.org/10.1007/s00253-012-4550-x |
| حقوق: | CLOSED |
| رقم الأكسشن: | edsair.doi.dedup.....7a39bef88844a038f316250f409ffc60 |
| قاعدة البيانات: | OpenAIRE |
Find this article in full text from Springer Verlag. | تدمد: | 14320614 01757598 |
|---|