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  • 1
    Publication Date: 1999-08-07
    Description: The functions of many open reading frames (ORFs) identified in genome-sequencing projects are unknown. New, whole-genome approaches are required to systematically determine their function. A total of 6925 Saccharomyces cerevisiae strains were constructed, by a high-throughput strategy, each with a precise deletion of one of 2026 ORFs (more than one-third of the ORFs in the genome). Of the deleted ORFs, 17 percent were essential for viability in rich medium. The phenotypes of more than 500 deletion strains were assayed in parallel. Of the deletion strains, 40 percent showed quantitative growth defects in either rich or minimal medium.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Winzeler, E A -- Shoemaker, D D -- Astromoff, A -- Liang, H -- Anderson, K -- Andre, B -- Bangham, R -- Benito, R -- Boeke, J D -- Bussey, H -- Chu, A M -- Connelly, C -- Davis, K -- Dietrich, F -- Dow, S W -- El Bakkoury, M -- Foury, F -- Friend, S H -- Gentalen, E -- Giaever, G -- Hegemann, J H -- Jones, T -- Laub, M -- Liao, H -- Liebundguth, N -- Lockhart, D J -- Lucau-Danila, A -- Lussier, M -- M'Rabet, N -- Menard, P -- Mittmann, M -- Pai, C -- Rebischung, C -- Revuelta, J L -- Riles, L -- Roberts, C J -- Ross-MacDonald, P -- Scherens, B -- Snyder, M -- Sookhai-Mahadeo, S -- Storms, R K -- Veronneau, S -- Voet, M -- Volckaert, G -- Ward, T R -- Wysocki, R -- Yen, G S -- Yu, K -- Zimmermann, K -- Philippsen, P -- Johnston, M -- Davis, R W -- HG00185-02/HG/NHGRI NIH HHS/ -- HG01627/HG/NHGRI NIH HHS/ -- HG01633/HG/NHGRI NIH HHS/ -- etc. -- New York, N.Y. -- Science. 1999 Aug 6;285(5429):901-6.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Biochemistry, Stanford University School of Medicine, Stanford, CA 94305-5307, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/10436161" target="_blank"〉PubMed〈/a〉
    Keywords: Culture Media ; *Gene Deletion ; Gene Expression Regulation, Fungal ; Gene Targeting ; *Genes, Essential ; Genes, Fungal ; *Genome, Fungal ; *Open Reading Frames ; Phenotype ; Polymerase Chain Reaction ; Recombination, Genetic ; Saccharomyces cerevisiae/*genetics/growth & development
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 2
    Publication Date: 2011-09-16
    Description: Recent advances in DNA synthesis technology have enabled the construction of novel genetic pathways and genomic elements, furthering our understanding of system-level phenomena. The ability to synthesize large segments of DNA allows the engineering of pathways and genomes according to arbitrary sets of design principles. Here we describe a synthetic yeast genome project, Sc2.0, and the first partially synthetic eukaryotic chromosomes, Saccharomyces cerevisiae chromosome synIXR, and semi-synVIL. We defined three design principles for a synthetic genome as follows: first, it should result in a (near) wild-type phenotype and fitness; second, it should lack destabilizing elements such as tRNA genes or transposons; and third, it should have genetic flexibility to facilitate future studies. The synthetic genome features several systemic modifications complying with the design principles, including an inducible evolution system, SCRaMbLE (synthetic chromosome rearrangement and modification by loxP-mediated evolution). We show the utility of SCRaMbLE as a novel method of combinatorial mutagenesis, capable of generating complex genotypes and a broad variety of phenotypes. When complete, the fully synthetic genome will allow massive restructuring of the yeast genome, and may open the door to a new type of combinatorial genetics based entirely on variations in gene content and copy number.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3774833/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉   〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3774833/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Dymond, Jessica S -- Richardson, Sarah M -- Coombes, Candice E -- Babatz, Timothy -- Muller, Heloise -- Annaluru, Narayana -- Blake, William J -- Schwerzmann, Joy W -- Dai, Junbiao -- Lindstrom, Derek L -- Boeke, Annabel C -- Gottschling, Daniel E -- Chandrasegaran, Srinivasan -- Bader, Joel S -- Boeke, Jef D -- AG023779/AG/NIA NIH HHS/ -- R01 AG023779/AG/NIA NIH HHS/ -- R37 AG023779/AG/NIA NIH HHS/ -- England -- Nature. 2011 Sep 14;477(7365):471-6. doi: 10.1038/nature10403.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉High Throughput Biology Center, Johns Hopkins University School of Medicine, 733 North Broadway, Baltimore, Maryland 21205, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/21918511" target="_blank"〉PubMed〈/a〉
    Keywords: Attachment Sites, Microbiological/genetics ; Chromosomes, Artificial, Yeast/*genetics ; Directed Molecular Evolution/methods ; Gene Dosage/genetics ; Gene Expression Profiling ; Gene Expression Regulation, Fungal ; Genetic Engineering/*methods ; Genetic Fitness/genetics ; Genome, Fungal/genetics ; Genotype ; Haploidy ; Molecular Sequence Data ; Mutagenesis/genetics ; Phenotype ; RNA, Fungal/analysis/genetics ; Saccharomyces cerevisiae/classification/*genetics ; Synthetic Biology/*methods
    Print ISSN: 0028-0836
    Electronic ISSN: 1476-4687
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
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