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J Virol. 1988 October; 62(10): 3738-3746

Use of simian virus 40 replication to amplify Epstein-Barr virus shuttle vectors in human cells.

S S Heinzel, P J Krysan, M P Calos and R B DuBridge

Department of Genetics, Stanford University School of Medicine, California 94305.

ABSTRACT

We have increased the copy number of Epstein-Barr virus vectors that also carry the origin of replication of simian virus 40 (SV40) by providing a transient dose of SV40 T antigen. T antigen was supplied in trans by transfection of a nonreplicating plasmid which expresses T antigen into cells carrying Epstein-Barr virus-SV40 vectors. A significant increase in vector copy number occurred over the next few days. We also observed a high frequency of intramolecular recombination when the vector carried a repeat segment in direct orientation, but not when the repeat was in inverted orientation or absent. Furthermore, by following the mutation frequency for a marker on the vector after induction of SV40 replication, it was determined that SV40 replication generates a detectable increase in the deletion frequency but no measurable increase in the frequency of point mutations.


J Virol. 1988 October; 62(10): 3738-3746




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