Global ETD Search

1	Metagenomic discovery and characterisation of restriction endonuclease from Kogelberg Biosphere Reserve Mtimka, Sibongile 05 1900 (has links) Restriction endonucleases are a group of enzymes that cleave DNA at or around specific sequences, which are typically palindromic. A fosmid library was constructed from a metagenome isolated from soil from the Kogelberg Nature Reserve, Western Cape and was functionally screened for restriction endonucleases. Next-generation (NGS) Illumina sequencing technology was used to identify putative endonucleases. The sequence data generated was assembled and analysed using CLC Bio Genomics Workbench and bioinformatics tools (NCBI BLAST, REBASE and MG-RAST). Using these tools, genes encoding restriction-modification systems and endonuclease homologues were discovered. Three genes were identified and were recombinantly produced in Rosetta™ (DE3) pLysS and purified with IMAC using Ni-TED resin and subsequently characterised. These three genes were selected based on the identity percentage when compared to sequences on the NCBI database. Production of Endo8 was scaled up using 2 l fermenter and the purification done using ÄKTA Avant 150 FPLC using a HiScale 50 column packed with Ni-TED resin and the total amount of protein achieved was 58.82 mg.g-1. The productivity achieved at 17 hours (8 h harvest) was 2-fold greater than at 12 hours. Endonuclease activity of endo8 and endo52 was tested, both exhibited strong non-specific activity at 37 °C with an incubation period of 30 min. This work demonstrates that environmental soil samples are a valuable source for discovery of novel enzymes and also the utility of functional metagenomics to discover and purify these enzymes. These endonucleases may contribute to the next generation of reagent enzymes for molecular biology research. / Chemistry / M. Sc. (Life Sciences) Bacteriophage Fosmid library Functional screening Kogelberg Nature Reserve Restriction endonuclease Restriction enzymes Soil metagenome 572.786 Restriction enzymes, DNA Bacteriophages -- Genetics Endonucleases
2	Identification and Characterization of Microbial Key Functions in Soils of the German Biodiversity Exploratories Representing Different Land Use and Management Types / Identifizierung und Charakterisierung von mikrobiellen Schlüsselfunktionen in Böden unterschiedlichen Landnutzungs- und Managementtyps der deutschen Biodiversitäts-Exploratorien Nacke, Heiko 20 October 2011 (has links) No description available. 570 Biowissenschaften, Biologie WUV 000 WUK 000 Biology (incl. Psychology) Bodenmetagenom Pyrosequenzierung bakterielle Diversität metagenomische Genbibliotheken lipolytische Enzyme (hemi)cellulolytische Enzyme Soil metagenome pyrosequencing bacterial diversity metagenomic libraries lipolytic enzymes (hemi)cellulolytic enzymes 42.30 42.13
3	Identifikation von Genen und Mikroorganismen, die an der dissimilatorischen Fe(III)-Reduktion beteiligt sind / Isolation of Genes and Microorganisms Involved in Dissimilatory Fe(III)-Reduction Özyurt, Baris 21 January 2009 (has links) No description available. 500 Naturwissenschaften allgemein Mathematics and Computer Science Metagenomik dissimilatorische Fe(III)-Reduktion Dissimilatory iron reduction DIR dissimilatory oxido-reduction of iron dissimilatory iron(III) reduction metal reduction ferricironreduction microbial iron respiration bacterial respiration microorganisms for energy production dissimilatory iron reducing bacteria dissimilatory metal reducing bacteria DMRB energy production electricity energy transduction microbial electricity electron donor electron acceptor electron transfer carbon source carbohydrate mannit mannitol biofilm mineral mineral formation microbial degradation ribosomal proteins cytochromes hypothetical proteins enzymes oxidation reduction ferric iron iron oxide Fe(II) Fe(III) soluble iron insoluble iron ferric reductase activity iron mineral formation mineralisation <i>E. coli</i> Escherichia coli environmental microorganisms life in extreme environments uncultivable bacteria soil microbial ecology anaerobic growth anaerobic incubation environmental sequences environmental DNA DNA extraction functional screening metagenomic metagenome meta-genome microbial ecology environmental genomics soil metagenome BACs sequencing bioinformatics microbial communities 16S rRNA phylogenetics 30 42.30 42.30 58.30 42.49 58.30 42.13 RA W

Search results

Metagenomic discovery and characterisation of restriction endonuclease from Kogelberg Biosphere Reserve

Identifikation von Genen und Mikroorganismen, die an der dissimilatorischen Fe(III)-Reduktion beteiligt sind / Isolation of Genes and Microorganisms Involved in Dissimilatory Fe(III)-Reduction