Characterisation of a DNA ligase from an Antarctic metagenomic library

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dc.contributor.advisorCowan, D. A
dc.contributor.authorBooysen, Dean
dc.date.accessioned2015-02-26T13:00:54Z
dc.date.accessioned2024-05-09T07:45:20Z
dc.date.available2015-02-26T13:00:54Z
dc.date.available2024-05-09T07:45:20Z
dc.date.issued2011
dc.description>Magister Scientiae - MScen_US
dc.description.abstractA metagenomic gene library prepared from soil found beneath a mummified seal carcass in the Miers Valley, Antarctica, suggests an environment rich in uncharacterised biodiversity including enzymes with possible application to industrial processes. A sequence based gene mining investigation was performed on a clone, which archives a metagenomic sequence from this environment. The sequence was annotated using de novo bioinformatics and molecular biology techniques. A predicted NAD+-dependent DNA ligase, ligDB1 was selected for further characterisation. LigDB1 encodes a gene product that contains all the sequence features of a functional ligase. The protein was overexpressed in a heterologous E. coli host and purified to homogeneity. LigDB1 did not exhibit nick sealing activity, but was able to perform AMP-dependent DNA relaxation in the presence of high concentrations of enzyme. DNA modifying enzymes from cold environments perform optimally at low temperatures and may be of use as molecular tools in biotechnology. Complete characterisation of this enzyme is subject to further investigationsen_US
dc.identifier.urihttps://hdl.handle.net/10566/13318
dc.language.isoenen_US
dc.publisherUniversity of the Western Capeen_US
dc.rights.holderUniversity of the Western Capeen_US
dc.subjectMetagenomic DNAen_US
dc.subjectsequence based screeningen_US
dc.subjectNAD+-dependent DNA ligaseen_US
dc.subjectligaseen_US
dc.subjectAntarcticaen_US
dc.subjectbioinformaticsen_US
dc.titleCharacterisation of a DNA ligase from an Antarctic metagenomic libraryen_US
dc.typeThesisen_US

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