Construction of rat cell lines that contain potential morphologically transforming regions of the herpes simplex virus type 2 genome

• Published in: Intervirology Vol. 24; no. 4; p. 199
• Main Authors: van den Berg, F M, van Amstel, P J, Walboomers, J M
• Format: Journal Article
• Language: English
• Published: Switzerland 01.01.1985
• Subjects: Animals; Base Sequence; Cell Division; Cell Line; Cell Transformation, Neoplastic; Cell Transformation, Viral; Cloning, Molecular; DNA, Viral - genetics; Genes, Viral; Nucleic Acid Hybridization; Plasmids; Rats; Simplexvirus - enzymology; Simplexvirus - genetics; Thymidine Kinase - genetics; Transcription, Genetic; Transfection
• ISSN: 0300-5526
• DOI: 10.1159/000149644

Hybrid recombinant plasmids were constructed; they were composed of the herpes simplex virus type 2 (HSV2) thymidine kinase (tk) gene and DNA sequences of HSV2 that have been reported to induce morphological and/or oncogenic transformation of rodent cells in culture. Several plasmids were made in two versions, with or without the simian virus 40 enhancer sequences. These plasmids were employed to transfect contact-inhibited Rat-2 tk- cells. It was demonstrated that cell lines with stably integrated, morphologically transforming regions (MTRs) of HSV2 could be isolated at a low frequency. In addition, many cell lines were isolated that had lost the MTR moiety of the transforming plasmids. None of the isolated tk-positive cell lines exhibited significantly altered growth properties when compared to control cell lines. Transient expression of transfecting enhancer plasmids was detectable 40 h posttransfection, but no transformed cell lines were obtained from these experiments. This conflicted with some versions of the 'hit-and-run' hypothesis for HSV-mediated cell transformation.