Detection of virus-specific DNA sequences by quantitative in situ hybridization during infection with SV40

✍ Scribed by Yves Langelier; Rosemonde Mandeville; René Simard

• Publisher: John Wiley and Sons
• Year: 1975
• Tongue: French
• Weight: 880 KB
• Volume: 16
• Category: Article
• ISSN: 0020-7136
• DOI: 10.1002/ijc.2910160315

✦ Synopsis

Abstract

In situ hybridization has been used as a quantitative method to study the replication of SV40 during lytic infection. Various parameters have been defined such as fixation and denaturation of cytological preparations, concentration of complementary (^3^H)‐cRNA and volume of buffer used for the hybridization reaction. When all these parameters are carefully controlled, a reproducibility of ±10% can be obtained for the quantitative study of the system SV40‐monkey kidney cells (CV‐1). In this system, it is evident that the synthesis of viral DNA is not localized in a special area of the nucleus and that it is not synchronized in confluent cells. Moreover, a proportion of 20 to 30% of cells are not labelled whatever the multiplicity of infection. In SV40‐transformed cells, it was not possible to detect significantly the integrated genome due either to the small size of the genome or to the fact that the number of genome‐equivalents integrated at the same place is too low to yield a radioactivity superior to background level for long periods of exposure.