1887

Abstract

We previously described the insertion of a foreign gene into a non-essential region of human cytomegalovirus (HCMV) by homologous recombination. Here we report insertion of the gene downstream of the mouse metallothionein promoter into the dIII-O region of HCMV by replacement-type recombination. Expression of the gene in the recombinant was independent of viral growth, but dependent on induction by heavy metals. Of several metals tested for -galactosidase induction and also for their toxicity to HEL cells, Zn was found to be the most suitable for use as an inducer. In HEL cells infected with the recombinant in the presence of 50 μ-Zn, -galactosidase activity was maximal 3 days after infection, and reached levels 27 times higher than the value obtained in the absence of Zn.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-74-8-1649
1993-08-01
2022-08-16
Loading full text...

Full text loading...

/deliver/fulltext/jgv/74/8/JV0740081649.html?itemId=/content/journal/jgv/10.1099/0022-1317-74-8-1649&mimeType=html&fmt=ahah

References

  1. Chen C, Okayama H. 1987; High efficiency transformation of mammalian cells by plasmid DNA.. Molecular and Cellular Biology 7:2745–2752
    [Google Scholar]
  2. Durnam DM, Palmiter RD. 1981; Transcriptional regulation of the mouse metallothionein-I gene by heavy metals.. Journal of Biological Chemistry 256:5712–5716
    [Google Scholar]
  3. Durnam DM, Perrin F, Gannon F, Palmiter RD. 1980; Isolation and characterization of the mouse metallothionein-I gene.. Proceedings of the National Academy of Sciences, U.S.A. 77:6511–6515
    [Google Scholar]
  4. Ihara S, Takekoshi M, Watanabe Y. 1986; Cleavage map of human cytomegalovirus genome (strain Towne) determined by the use of cosmid cloning system.. Archives of Virology 88:241–250
    [Google Scholar]
  5. Johnson PA, Yoshida K, Gage FH, Friedmann T. 1992; Effects of gene transfer into cultured CNS neurons with a replication-defective herpes simplex virus type 1 vector.. Molecular Brain Research 12:95–102
    [Google Scholar]
  6. Jones TR, Muzithras VP, Gluzman Y. 1991; Replacement mutagenesis of the human cytomegalovirus genome: US 10 and US11 gene products are nonessential.. Journal of Virology 65:5860–5872
    [Google Scholar]
  7. Lowe RSP, Keller M, Keech BJ, Davison AJ, Whang Y, Morgan AJ, Kieff E, Ellis RW. 1987; Varicella-zoster virus as a live vector for the expression of foreign genes.. Proceedings of the National Academy of Sciences, U.S.A. 84:3896–3900
    [Google Scholar]
  8. Mackett M, Smith GL. 1986; Vaccinia virus expression vectors.. Journal of General Virology 67:2067–2082
    [Google Scholar]
  9. Manning WC, Mocarski ES. 1988; Insertion mutagenesis of the murine cytomegalovirus genome: one prominent a gene (ie2) is dispensable for growth.. Virology 167:477–484
    [Google Scholar]
  10. Miller JH. 1972; Assay of β-galactosidase.. In Experiments in Molecular Genetic pp 398–104 New York: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  11. Mocarski ES, Post LE, Roizman B. 1980; Molecular engineering of the herpes simplex virus genome: insertion of a second L-S junction into the genome causes additional genome inversion.. Cell 22:243–255
    [Google Scholar]
  12. Puckett C, Moss B. 1983; Selective transcription of vaccinia virus genes in template dependent soluble extracts of infected cells.. Cell 35:441–448
    [Google Scholar]
  13. Roizman B, Jenkins FJ. 1985; Genetic engineering of novel genomes of large DNA viruses.. Science 229:1208–1214
    [Google Scholar]
  14. Shirakawa M, Tsurimoto T, Matsubara K. 1984; Plasmid vectors designed for high-efficiency expression controlled by the portable recA promoter-operator of Escherichia coli.. Gene 28:127–132
    [Google Scholar]
  15. Spate RR, Mocarski ES. 1987; Insertion and deletion mutagenesis of the human cytomegalovirus genome.. Proceedings of the National Academy of Sciences, U.S.A. 84:7213–7217
    [Google Scholar]
  16. Tackney G, Cachianes G, Silverstein S. 1984; Transduction of the Chinese hamster ovary aprt gene by herpes simplex virus.. Journal of Virology 52:606–614
    [Google Scholar]
  17. Takekoshi M, Maeda-Takekoshi F, Ihara S, Sakuma S, Watanabe Y. 1991; Site-specific stable insertion into the human cytomegalovirus genome of a foreign gene under control of the SV40 promoter.. Gene 101:209–213
    [Google Scholar]
  18. Thomsen DR, Marotti KR, Palermo DP, Post LE. 1987; Pseudorabies virus as a live virus vector for expression of foreign genes.. Gene 57:261–265
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-74-8-1649
Loading
/content/journal/jgv/10.1099/0022-1317-74-8-1649
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error