GENE EXPRESSION PROFILES IN VIRUS PRODUCING AND LOW/NON VIRUS PRODUCING EBV-BAC CONTAINING CELL LINES

摘要

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 line's 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.