Infection with herpes simplex virus leads to amplification of SV40 DNA in various SV40-transformed cells. In earlier studies with the SV40-transformed hamster cell line Elona two different types of DNA amplification could be identified: (i) Bidirectional overreplication of chromosomally integrated SV40 DNA expanding into the flanking cellular sequences ("onion skin" type) and (ii) highly efficient synthesis of extremely large head-to-tail concatemers containing exclusively SV40 DNA ("rolling circle" type). These investigations have indicated that the chromosomally integrated form of SV40 might be the substrate for both types of overreplication. There still had been uncertainties as to whether and how these events were connected. A hypothetical assumption of a recombinational event leading to the excision of SV40 DNA molecules is supported by the results presented here: In this study cloned Elona cell lines were investigated for their ability to amplify SV40 sequences and for the mechanism of amplification utilized. SV40 integration in a partial tandem manner correlates with a strong rolling circle amplification. In contrast, in one cell line harboring a truncated SV40 genome, amplification appears mainly restricted to intrachromosomal bidirectional overreplication. Possible implications for HSV functions involved in the amplification process will be discussed.