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Title: | Construction of an Adenovirus Expression Vector Containing the T4 Den V Gene, Which Can Complement the DNA Repair Deficiency of Xeroderma Pigmentosum Fibroblasts |
Other Titles: | Construction of an AD 5 Vector Containing the T4 Den V Gene |
Authors: | Colicos, Michael, A. |
Advisor: | Rainbow, Andrew, J. |
Department: | Biology |
Keywords: | adenovirus expression vector, T4 den v gene, DNA repair, DNA repair deficiency, zeroderma pigmentosum fibroblasts, xeroderma, pigmentosum fibroblasts, ad 5 vector |
Publication Date: | Aug-1988 |
Abstract: | This study demonstrates the use of an adenovirus vector system to study the effect of a DNA repair gene on untransformed human fibroblasts. The bacteriophage T4 pyrimidine dimer DNA glycosylase (den V) gene has been inserted into the E3 region of human adenovirus type 5. The resulting recombinant virus Ad Den V was determined to be producing correctly initiated RNA from the RSV 3' LTR promoter used in the den V expression cartridge inserted into the virus. The effect of the den V gene product on human fibroblasts 'liras examined by assaying for the percent host cell reactivation (%HCR) of Vag production for UV irradiated Ad Den V in comparison to that for a control virus. It was shown that the %HCR was significantly greater for Ad Den V as compared to the control virus in xeroderma pigmentosum (XP) cells. UV survival of adenovirus in XP cells exhibited a two component nature. Introduction of the den V gene into XP group A cells increased the D0 value of the first component of the viral survival curve to a level similar to that of XPC cells, which showed no change in this component irrespective of the presence of the den V gene. It has been suggested that the den V gene is able to partially complement the deficiency in some XP cells because of its small size, allowing it to gain access to the DNA damage site where as the cellular repair enzyme complex can not. Since XPC cells are proficient in their alteration of DNA secondary structure prior to DNA excision repair, these results are consistant with the hypothesis that the first component of UV viral survival curves reflects the pathway involved in accessing the damaged sites. The manuscript of a paper has been included as an appendix. The work theorizes on the origin of mammalian immune system diversity and bacteriophage lambda, and their possible relationship to prokaryotic DNA repair genes. |
URI: | http://hdl.handle.net/11375/23624 |
Appears in Collections: | Digitized Open Access Dissertations and Theses |
Files in This Item:
File | Description | Size | Format | |
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Colicos_Michael_A_Aug1988_masters0001-.pdf | 17.27 MB | Adobe PDF | View/Open |
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