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The establishment of two novel bovine cell lines by transfection with the putative transforming region of bovine adenovirus type 3

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Show simple item record White, Donald E. en_US 2009-07-09T18:49:35Z 2009-07-09T18:49:35Z 1996-07-09T18:49:35Z
dc.description.abstract Human adenoviruses (Ads), members of the family adenoviridae, are medium-sized DNA viruses which have been used as valuable research tools for the study of RNA processing, oncogenic transformation, and for the development of viral vectors for use in gene delivery and immunization technology. The left 12% of the linear Ad genollle codes for products which are necessary for the efficient replication of the virus, as well as being responsible for the forlllation of tumors in animallllodels. The establishlllent of the 293 cell line, by immortalization of human embryonic kidney cells with th~ E1 region of Ad type S (AdS), has facilitated extensive manipulation of the Ads and the development of recombinant Ad vectors. The study of bovine adenoviruses (BAVs), which cause mild respiratory and gastrointestinal infections in cattle has, on the other hand, been limited primarily to that of infectivity, immunology and clinicallllanifestations. As a result, any potential as gene delivery vehicles has not yet been realized. Continued research into the molecular biolo~gy of BAVs and the development of recolllbinant vectors would benefit from the development of a cell line analogous to that of the 293 cells. In an attelllpt to establish such a cell line, the recombinant plaslllid pKC-neo was constructed, containing the left 0-19.7% of the BAV type 3 (BAV3) genome, and the selectable marker for resistance to the aminoglycoside G418, a neomycin derivative. The plasmid construct was then used to transfect both the Madin-Darby bovine kidney (MDBK) -iicell line and primary bovine lung cells, after which G418-resistant foci were selected for analysis. Two cell lines, E61 (MDBK) and E24 (primary lung), were subsequently selected and analysed for DNA content, revealing the presence of the pKC-neo sequences in their respective genomes. In addition, BAV3 RNA transcripts were detected in the E61 cells. Although the presence of E1 products has yet to be confirmed in both cell lines, the E24 cells exhibit a phenotype characteristic of partial transformation by E1. The apparent immortalization of the primary lung cells will permit exploitation of their ability to take up exogenous DNA at high efficiency. en_US
dc.language.iso eng en_US
dc.publisher Brock University en_US
dc.subject Adenoviruses. en_US
dc.subject Mutation (Biology) en_US
dc.subject Cattle--Genetic engineering. en_US
dc.title The establishment of two novel bovine cell lines by transfection with the putative transforming region of bovine adenovirus type 3 en_US
dc.type Electronic Thesis or Dissertation en_US M.Sc. Biological Sciences en_US Masters en_US
dc.contributor.department Department of Biological Sciences en_US Faculty of Mathematics and Science en_US

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