Overcoming interference to retroviral superinfection results in amplified expression and transmission of cloned genes

R. K. Bestwick, S. L. Kozak, David Kabat

Research output: Contribution to journalArticle

81 Citations (Scopus)

Abstract

A procedure is described for stably expressing cloned genes at high levels in vertebrate cells and for obtaining these genes in high-titer virus preparations. The process uses retroviral vectors and mixtures of two 'packaging cell lines' that incorporate retroviral genomes into virions with different host-range envelopes. In these cocultures, interference barriers to superinfection are overcome, retroviral vectors can replicate in the absence of a transmissible helper virus, and the cells become infected with multiple copies of the provirus that contains the cloned gene. This procedure was used to amplify expression of the membrane glycoprotein that is encoded by Friend spleen focus-forming virus, a retrovirus that is replication defective in other cell cultures. Amplifications were measured at the DNA provirus, RNA, and protein levels. In addition, the human growth hormone gene was inserted into retroviral vectors and we observed amplifications of growth hormone synthesis and secretion. The amplified growth hormone was properly processed as indicated by immunoblot analyses. A vector is described (pSFF) that is exceptionally active in coculture amplification.

Original languageEnglish (US)
Pages (from-to)5404-5408
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume85
Issue number15
StatePublished - 1988

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Superinfection
Proviruses
Coculture Techniques
Growth Hormone
Genes
Spleen Focus-Forming Viruses
Helper Viruses
Human Growth Hormone
Host Specificity
Membrane Glycoproteins
Product Packaging
Retroviridae
Helper-Inducer T-Lymphocytes
Viral Load
Virion
Vertebrates
Cell Culture Techniques
Genome
RNA
Cell Line

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Overcoming interference to retroviral superinfection results in amplified expression and transmission of cloned genes. / Bestwick, R. K.; Kozak, S. L.; Kabat, David.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 85, No. 15, 1988, p. 5404-5408.

Research output: Contribution to journalArticle

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