The herpes simplex virus amplicon: A new eucaryotic defective-virus cloning-amplifying vector

Richard R. Spaete*, Niza Frenkel

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

380 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)295-304
Number of pages10
JournalCell
Volume30
Issue number1
DOIs
StatePublished - Aug 1982
Externally publishedYes

Funding

FundersFunder number
U. S. Public Health Service
National Science Foundation
National Cancer Institute
U.S. Public Health Service

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