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Citation link: http://nbn-resolving.de/urn:nbn:de:gbv:46-diss000119865
00011986.pdf
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Expression und biochemische und strukturelle Charakterisierung der RNA-abhängigen RNA-Polymerase des Lassa-Virus


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Other Titles: Expression and biochemical and structural characterisation of the RNA dependent RNA polymerase of Lassa virus
Authors: Brunotte, Linda 
Supervisor: Dotzauer, Andreas
1. Expert: Dotzauer, Andreas
2. Expert: Guenther, Stephan
Abstract: 
In this work the domain structure of the Lassa virus L protein was investigated. Potential inter-domainlinkers were predicted and functionally analyzed in the minireplicon system. Two sites were identified at which L protein could be physically separated without significant loss of function in the minireplicon system, indicating that the corresponding domains are able to trans-complement each other. Physical interaction of the N- and C-terminal domains was verified by co-immunoprecipitation and confocal immunofluorescence microscopy. The data indicate that the L protein is composed of at least three domains.In the second part full length L proteins of Lassa virus AV, Bantou and Mopeia virus should be expressed in three different expression systems. Furthermore, a polymerase assay should be established to allow biochemical characterisation of the polymerase activity of L protein. Expression of L protein was successfull in insect cells and HeLa cells, but not bacteria. The rekombinant proteins could be purified using either His- or Strep-Tag but it was not possible to detect polymerase activity.
Keywords: Lassa virus, RNA dependent RNA polymerase
Issue Date: 16-Jun-2010
Type: Dissertation
URN: urn:nbn:de:gbv:46-diss000119865
Institution: Universität Bremen 
Faculty: FB2 Biologie/Chemie 
Appears in Collections:Dissertationen

  

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