Generalized Schemes for High-Throughput Manipulation of the Desulfovibrio vulgaris Genome
| العنوان: | Generalized Schemes for High-Throughput Manipulation of the Desulfovibrio vulgaris Genome |
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| المؤلفون: | Evelin D. Szakal, Simon Allen, Manfred Auer, Swapnil R. Chhabra, Mario Ouellet, Gareth Butland, Judy D. Wall, Jay D. Keasling, Harry R. Beller, Barbara Gold, Mark D. Biggin, R. Prathapam, H. E. Witkowska, Thomas R. Juba, Sonia A. Reveco, John-Marc Chandonia, Amita Gorur, Kimberly L. Keller, C. M. Leung, Terry C. Hazen, O.-Y. Fok, Danielle M. Jorgens, Grant M. Zane, Elizabeth S. Semkiw, Adam P. Arkin, Morgan N. Price, Mary E. Singer, Dwayne A. Elias |
| المصدر: | Applied and Environmental Microbiology. 77:7595-7604 |
| بيانات النشر: | American Society for Microbiology, 2011. |
| سنة النشر: | 2011 |
| مصطلحات موضوعية: | DNA, Bacterial, Genetics, Microbial, Genetic Vectors, Genomics, Computational biology, Applied Microbiology and Biotechnology, Genome, chemistry.chemical_compound, Methods, Desulfovibrio vulgaris, Gene, Recombination, Genetic, Genetics, Ecology, biology, Artificial Gene Fusion, biology.organism_classification, Protein subcellular localization prediction, Desulfovibrio, High-Throughput Screening Assays, Mutagenesis, Insertional, chemistry, Gene Deletion, Genome, Bacterial, DNA, Food Science, Biotechnology |
| الوصف: | The ability to conduct advanced functional genomic studies of the thousands of sequenced bacteria has been hampered by the lack of available tools for making high-throughput chromosomal manipulations in a systematic manner that can be applied across diverse species. In this work, we highlight the use of synthetic biological tools to assemble custom suicide vectors with reusable and interchangeable DNA “parts” to facilitate chromosomal modification at designated loci. These constructs enable an array of downstream applications, including gene replacement and the creation of gene fusions with affinity purification or localization tags. We employed this approach to engineer chromosomal modifications in a bacterium that has previously proven difficult to manipulate genetically, Desulfovibrio vulgaris Hildenborough, to generate a library of over 700 strains. Furthermore, we demonstrate how these modifications can be used for examining metabolic pathways, protein-protein interactions, and protein localization. The ubiquity of suicide constructs in gene replacement throughout biology suggests that this approach can be applied to engineer a broad range of species for a diverse array of systems biological applications and is amenable to high-throughput implementation. |
| تدمد: | 1098-5336 0099-2240 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::644e9a3a972cb8bbb93431b3fe1e95e1Test https://doi.org/10.1128/aem.05495-11Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....644e9a3a972cb8bbb93431b3fe1e95e1 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10985336 00992240 |
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