Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.796445
Title: In vitro transformation of non-established bovine fibroblasts by BPV4 and cooperating factors
Author: Jaggar, Rhys Trevor
Awarding Body: University of Glasgow
Current Institution: University of Glasgow
Date of Award: 1990
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Abstract:
An in vitro assay system utilising non-established bovine fibroblasts derived from foetal palate was developed for the detection of BPV4-encoded transforming functions. This involved cotransfection of a dominant coselectable marker gene neo and recombinant BPV4 DNA linearised within the E1 ORF. The presence of BPV4 DNA resulted in the formation of colonies of diameter greater than 5mm (macrocolonies) which displayed a contact-inhibited phenotype. Cloning of regions of BPV4 into two vectors, pSV2neo and pZIPneoSV(X1), indicated that macrocolony formation could be induced by two regions of the genorpe. The first, a 2.0kb Xholl fragment (nts 6487-1275) encoded two complete ORFs, namely E7 and E8. A construct lacking 233bp (nts 906-1139), resulting in interruption of the E7 ORF, was inactive in this assay, indicating a requirement for the E7 ORF for induction of the phenotype. The second region, encoded by a 3.9kb Xholl fragment (nts 2597-6487), contained the E2, E3, E4, E5 and L2 ORFs. In this case, colonies contained cells showing a more elongated phenotype. Cotransfections of linear BPV4 with an activated ras gene did not lead to a loss of contact-inhibition. In contrast, overexpression of the 2.0kb fragment, in the presence of ras did lead to the formation of non-contact-inhibited neo r macrocolonies. An intact E7 ORF was again required for this activity. In the case of the 3.9kb construct, a single non-contact-inhibited colony was observed in constructs utilising the BPV4 promoters upon cotransfection with ras.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.796445  DOI: Not available
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