Use of simian virus 40 replication to amplify Epstein-Barr virus shuttle vectors in human cells.
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.
Heinzel
S S
SS
Department of Genetics, Stanford University School of Medicine, California 94305.
Krysan
P J
PJ
Calos
M P
MP
DuBridge
R B
RB
D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D002460:Cell Line; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis / Q000235:genetics; D003720:Densitometry; D005784:Gene Amplification; D005822:Genetic Vectors; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000502:physiology; D006801:Humans; D009154:Mutation; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D013539:Simian virus 40 / Q000235:genetics* / Q000502:physiology; D014162:Transfection; D014779:Virus Replication
D000952:Antigens, Polyomavirus Transforming; D004279:DNA, Viral