Conditional expression of foreign genes by temperature-sensitive mutants of vaccinia virus

Thomas M. Chambers; Karim Essani; Robert G. Webster

doi:10.1016/0378-1119(90)90371-W

Conditional expression of foreign genes by temperature-sensitive mutants of vaccinia virus

Thomas M. Chambers, Karim Essani, Robert G. Webster

Veterinary Science

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

To assess the utility of two temperature-sensitive (ts) mutant vaccinia viruses as vectors for the conditional in vitro expression of recombinant foreign genes, we have studied the kinetics of expression of foreign genes incorporated into these viruses. At nonpermissive temperature, 40°C, these viruses were defective either in DNA synthesis or in virus assembly. Foreign gene expression was affected by the nature of the ts lesion and by the nature of the vaccinia promoter positioned upstream from the foreign gene. With both vector viruses, a foreign gene controlled by the p7.5 early-late promoter was expressed at both 33° and 40°C. With the DNA synthesis-defective vector virus, foreign gene expression controlled by the p11 DNA synthesis-dependent late promoter was inhibited at 40°C, but could be turned on by shift to 33°C. This ts expression system provides an alternative to use of drugs that inhibit DNA synthesis as a means for experimental manipulation of gene expression. Both vector viruses can be used with existing vaccinia virus expression technology.

Original language	English
Pages (from-to)	275-278
Number of pages	4
Journal	Gene
Volume	95
Issue number	2
DOIs	https://doi.org/10.1016/0378-1119(90)90371-W
State	Published - Nov 15 1990

Bibliographical note

Funding Information:
This work was supported by USPHS Contract No. AI-52586 from the National Institutes of Health, and by American Lebanese Syrian Associated Charities. We thank Samuel Dales for providing the ts mutant vaccinia viruses, Bernard Moss for providing the pSCI 1 plasmid, and John M. Freeman for technical assistance.

Funding

This work was supported by USPHS Contract No. AI-52586 from the National Institutes of Health, and by American Lebanese Syrian Associated Charities. We thank Samuel Dales for providing the ts mutant vaccinia viruses, Bernard Moss for providing the pSCI 1 plasmid, and John M. Freeman for technical assistance.

Funders	Funder number
National Institutes of Health (NIH)
National Institute of Allergy and Infectious Diseases	N01AI052586
U.S. Public Health Service
American Lebanese Syrian Associated Charities

Keywords

DNA synthesis inhibition
Recombinant DNA
gene expression vector
influenza hemagglutinin
transcription promoter
β-galactosidase

ASJC Scopus subject areas

Genetics

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/0378-1119(90)90371-W

Cite this

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title = "Conditional expression of foreign genes by temperature-sensitive mutants of vaccinia virus",

abstract = "To assess the utility of two temperature-sensitive (ts) mutant vaccinia viruses as vectors for the conditional in vitro expression of recombinant foreign genes, we have studied the kinetics of expression of foreign genes incorporated into these viruses. At nonpermissive temperature, 40°C, these viruses were defective either in DNA synthesis or in virus assembly. Foreign gene expression was affected by the nature of the ts lesion and by the nature of the vaccinia promoter positioned upstream from the foreign gene. With both vector viruses, a foreign gene controlled by the p7.5 early-late promoter was expressed at both 33° and 40°C. With the DNA synthesis-defective vector virus, foreign gene expression controlled by the p11 DNA synthesis-dependent late promoter was inhibited at 40°C, but could be turned on by shift to 33°C. This ts expression system provides an alternative to use of drugs that inhibit DNA synthesis as a means for experimental manipulation of gene expression. Both vector viruses can be used with existing vaccinia virus expression technology.",

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author = "Chambers, {Thomas M.} and Karim Essani and Webster, {Robert G.}",

note = "Funding Information: This work was supported by USPHS Contract No. AI-52586 from the National Institutes of Health, and by American Lebanese Syrian Associated Charities. We thank Samuel Dales for providing the ts mutant vaccinia viruses, Bernard Moss for providing the pSCI 1 plasmid, and John M. Freeman for technical assistance.",

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N2 - To assess the utility of two temperature-sensitive (ts) mutant vaccinia viruses as vectors for the conditional in vitro expression of recombinant foreign genes, we have studied the kinetics of expression of foreign genes incorporated into these viruses. At nonpermissive temperature, 40°C, these viruses were defective either in DNA synthesis or in virus assembly. Foreign gene expression was affected by the nature of the ts lesion and by the nature of the vaccinia promoter positioned upstream from the foreign gene. With both vector viruses, a foreign gene controlled by the p7.5 early-late promoter was expressed at both 33° and 40°C. With the DNA synthesis-defective vector virus, foreign gene expression controlled by the p11 DNA synthesis-dependent late promoter was inhibited at 40°C, but could be turned on by shift to 33°C. This ts expression system provides an alternative to use of drugs that inhibit DNA synthesis as a means for experimental manipulation of gene expression. Both vector viruses can be used with existing vaccinia virus expression technology.

AB - To assess the utility of two temperature-sensitive (ts) mutant vaccinia viruses as vectors for the conditional in vitro expression of recombinant foreign genes, we have studied the kinetics of expression of foreign genes incorporated into these viruses. At nonpermissive temperature, 40°C, these viruses were defective either in DNA synthesis or in virus assembly. Foreign gene expression was affected by the nature of the ts lesion and by the nature of the vaccinia promoter positioned upstream from the foreign gene. With both vector viruses, a foreign gene controlled by the p7.5 early-late promoter was expressed at both 33° and 40°C. With the DNA synthesis-defective vector virus, foreign gene expression controlled by the p11 DNA synthesis-dependent late promoter was inhibited at 40°C, but could be turned on by shift to 33°C. This ts expression system provides an alternative to use of drugs that inhibit DNA synthesis as a means for experimental manipulation of gene expression. Both vector viruses can be used with existing vaccinia virus expression technology.

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Conditional expression of foreign genes by temperature-sensitive mutants of vaccinia virus

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

UN SDGs

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