Detection and quantitation of simian virus 40 genetic material in abortively transformed BALB-3T3 clones (mice-diploid cells-virus equivalents)

Abstract

Infection with simian virus 40 is known to induce many cells to synthesize DNA and to divide in a medium lacking serum protein growth factor(s) that is essential for growth of uninfected cells (factor-free medium). Cells infected under these conditions then go through several rounds of division, since colonies containing more than 100 cells are formed. Many of these colonies are abortively transformed since, upon subsequent passage of the cells in standard medium, they can no longer grow in factor-free medium and show no other properties of viral transformation. We have examined these abortively transformed cells for the presence of simian virus 40 DNA sequences. Of the three clones tested, two were found to contain viral genetic material despite the fact that they were phenotypically normal. The number of simian virus 40 genome equivalents present was determined by measurement of DNA reassociation kinetics on hydroxyapatite. Two of the abortively transformed lines contained approximately five viral genome equivalents per diploid cell, while the DNA from a third abortive transformant was indistinguishable from that of uninfected BALB/3T3 cells. A standard simian virus 40 transformant, isolated under similar conditions, contained two copies of the viral genome per cell. The abortive transformants also appear to contain the entire viral genome rather than multiple partial copies. Subclones of one abortively transformed line containing five copies per cell had 2.7-10 copies of viral genetic material per diploid cell.