TY - JOUR
T1 - The herpes simplex virus amplicon
T2 - A new eucaryotic defective-virus cloning-amplifying vector
AU - Spaete, Richard R.
AU - Frenkel, Niza
N1 - Funding Information:
We thank Ms. Glynis McCray and Betty Kwong for excellent technical assistance; Louis P. Deiss for most helpful discussions, and for derivation of the pP2-103 clone; and Dr. Byron Murray for the serially passaged Patton stocks. These studies were supported by grants from the U. S. Public Health Service, the National Cancer Institute and the National Science Foundation. R. R. S. is a predoctoral trainee supported by a Training Grant from the U.S. Public Health Service.
PY - 1982/8
Y1 - 1982/8
N2 - We have employed repeat units of herpes simplex virus (HSV) defective genomes to derive a cloning- amplifying vector (amplicon) that can replicate in eucaryotic cells in the presence of standard HSV helper virus. The design of the HSV amplicon system is based on the previous observation that cotransfection of cells with helper virus DNA and seed monomeric repeat units of HSV defective genomes results in the regeneration of concatemeric defective genomes composed of multiple reiterations of the seed repeats. Cotransfection of cells with helper virus DNA and chimeric repeat units containing bacterial plasmid pKC7 DNA resulted in the generation of defective genomes composed of reiterations of the seed HSV-pKC7 repeats. These chimeric defective genomes were packaged into virus particles and could be propagated in virus stocks, with the most enriched passages containing more than 90% chimeric defective genomes. Furthermore, monomeric chimeric repeat units could be transferred back and forth between bacteria and eucaryotic cells. A derivative vector constructed so as to contain several unique restriction enzyme sites could be potentially employed in the introduction of additional viral or eucaryotic DNA sequences into eucaryotic cells.
AB - We have employed repeat units of herpes simplex virus (HSV) defective genomes to derive a cloning- amplifying vector (amplicon) that can replicate in eucaryotic cells in the presence of standard HSV helper virus. The design of the HSV amplicon system is based on the previous observation that cotransfection of cells with helper virus DNA and seed monomeric repeat units of HSV defective genomes results in the regeneration of concatemeric defective genomes composed of multiple reiterations of the seed repeats. Cotransfection of cells with helper virus DNA and chimeric repeat units containing bacterial plasmid pKC7 DNA resulted in the generation of defective genomes composed of reiterations of the seed HSV-pKC7 repeats. These chimeric defective genomes were packaged into virus particles and could be propagated in virus stocks, with the most enriched passages containing more than 90% chimeric defective genomes. Furthermore, monomeric chimeric repeat units could be transferred back and forth between bacteria and eucaryotic cells. A derivative vector constructed so as to contain several unique restriction enzyme sites could be potentially employed in the introduction of additional viral or eucaryotic DNA sequences into eucaryotic cells.
UR - http://www.scopus.com/inward/record.url?scp=0020451027&partnerID=8YFLogxK
U2 - 10.1016/0092-8674(82)90035-6
DO - 10.1016/0092-8674(82)90035-6
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AN - SCOPUS:0020451027
SN - 0092-8674
VL - 30
SP - 295
EP - 304
JO - Cell
JF - Cell
IS - 1
ER -