Integration sites and sequence arrangement of SV40 DNA in a homogeneous series of transformed rat fibroblast lines.

Abstract:

:The state and organization of viral DNA sequences present in independently isolated rat cell lines transformed with SV40 were investigated using restriction-enzyme cleavage of the cellular DNA and blot hybridization with a viral probe. The transformed lines were established under conditions as identical as possible, except for a limited number of variables (multiplicity of infection, physiological state of the cells after infection and procedures used for selecting the transformed derivatives). They were characterized after a limited number of generations in culture. Two distinct types of organization were found: covalently integrated viral genomes were present either as single inserts or as head-to-tail oligomeric structures. The latter was observed among transformants derived from cells maintained after infection under growth-inhibiting conditions (suspension in agarose medium, confluency on a solid substrate). Single inserts were observed only among cell lines isolated after an initial period of active growth. Recurrent patterns of hybridizaton were observed in independently isolated lines, indicating that the sites of the integrative recombinations were close enough, both in the viral and the cellular sequences, not to be distinguished at the level of sensitivity of the technique (more than +/- 100 bp). Among cell lines with multiple integration sites, only part of the inserts were found in several instances to be identical to inserts observed in other transformed lines.

journal_name

Cell

journal_title

Cell

authors

Mougneau E,Birg F,Rassoulzadegan M,Cuzin F

doi

10.1016/0092-8674(80)90569-3

subject

Has Abstract

pub_date

1980-12-01 00:00:00

pages

917-27

issue

3

eissn

0092-8674

issn

1097-4172

pii

0092-8674(80)90569-3

journal_volume

22

pub_type

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