Our lab studies Epstein-Barr virus (EBV); therefore, we need a supply of cells that steadily produce virus for use in our experiments. Currently virus harvesting is unpredictable, as transfection of 293SL cells with a given Bacterial Artificial Chromosome (BAC) may produce cell lines that vary widely in the amount of infectious virus produced, with most lines producing no virus at all. Using quantitative real time PCR we quantified EBV mRNA expression pertaining to 16 specific targets including latent, lytic, and promoter transcripts. This was to determine if there was a correlation between the pattern of virus gene expression and the capacity of a cell line to produce virus. Such a correlation could be used to develop a screening assay predictive of a cell lines potential for virus production. We found that the genes most useful for creating a PCR-based screening assay were the genes belonging to the EBNA3 family. The public health significance of the steady production of Epstein-Barr virus would be that experiments could be conducted on quicker time tables, which in turn may increase the rate of knowledge obtained about EBV.
| Identifer | oai:union.ndltd.org:PITT/oai:PITTETD:etd-12072009-102318 |
| Date | 27 January 2010 |
| Creators | Lucas, Anna |
| Contributors | Dr. Jeremy Martinson, Dr. Frank Jenkins, Dr. David T. Rowe |
| Publisher | University of Pittsburgh |
| Source Sets | University of Pittsburgh |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.library.pitt.edu/ETD/available/etd-12072009-102318/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to University of Pittsburgh or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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