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Nucleic Acids Research, 1981, Vol. 9, No. 16 4111-4128
© 1981

MOLECULAR BIOLOGY

Characterization of a gene encoding a 115 K super T antigen expressed by a SV40-transformed rat cell line

Evelyne May*, Jean-Marc Jeltsch** and Frank Gannon**

*Institut de Recherches Scientifiques sur le Cancer B.P. no 8, 94800 Villejuif **Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS, Unité 184 de Biologie Moléculaire de Génie Génétique de l'INSERM Faculté de Médecine, Strasbourg, France

Received June 17, 1981. It has been reported that SV40-transformed V 11 F 1 clone 1 subclone 7 rat cells (subclone 7) produce a super T antigen of 115,000 M. This super T antigen is entirely SV40 coded and is synthesized by translation of an elongated form of SV40 early mRNA (May, E., Kress, M., Daya-Grosjean, L., Monier, R. and May, P. (1981) J. Virol., 37, 24–35). The results reported here show that there is only one independent insertion of viral DNA in the cellular genome of subclone 7 cells. When DNA from subclone 7 cells was cleaved with Bam HI endonuclease two distinct SV40 sequence containing fragments were generated with sizes of 5 Kb and 10 Kb, respectively.

Two recombinant cosmids were constructed by insertion of the 5 Kb and 10 Kb fragments, respectively, into cosmid pHC 79. Using restriction map analysis and nucleotide sequencing, we showed that the 5 Kb fragment actually contained the complete sequence of a gene encoding super T antigen. As compared to the normal SV40 early gene, the sequence of super T gene showed the following rearrangements : (i) The segment between nucleotides 4116 – 3544 was duplicated in a direct order and (ii) these two copies of 573 nucleotide sequence were separated by a 93 nucleotide tract which was a nearly perfect inverted repeat of the segment located between nucleotides 4868 and 4770 (nucleotide numbering used here = Weissmann number + 17).


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