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Neomycin resistance as a dominant selectable marker for selection and isolation of vaccinia virus recombinants

Franke, Christine A. and Rice, Charles M. and Strauss, James H. and Hruby, Dennis E. (1985) Neomycin resistance as a dominant selectable marker for selection and isolation of vaccinia virus recombinants. Molecular and Cellular Biology, 5 (8). pp. 1918-1924. ISSN 0270-7306. http://resolver.caltech.edu/CaltechAUTHORS:FRAmcb85

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Abstract

The antibiotic G418 was shown to be an effective inhibitor of vaccinia virus replication when an appropriate concentration of it was added to cell monolayers 48 h before infection. Genetic engineering techniques were used in concert with DNA transfection protocols to construct vaccinia virus recombinants containing the neomycin resistance gene (neo) from transposon Tn5. These recombinants contained the neo gene linked in either the correct or incorrect orientation relative to the vaccinia virus 7.5-kilodalton gene promoter which is expressed constitutively throughout the course of infection. The vaccinia virus recombinant containing the chimeric neo gene in the proper orientation was able to grow and form plaques in the presence of G418, whereas both the wild-type and the recombinant virus with the neo gene in the opposite polarity were inhibited by more than 98%. The effect of G418 on virus growth may be mediated at least in part by selective inhibition of the synthesis of a subset of late viral proteins. These results are discussed with reference to using this system, the conferral of resistance to G418 with neo as a positive selectable marker, to facilitate constructing vaccinia virus recombinants which contain foreign genes of interest.


Item Type:Article
Additional Information:Copyright © 1985 by the American Society for Microbiology. Received 19 February 1985/Accepted 9 May 1985 This research was supported by funds from the Murdoch Trust Foundation (to D.E.H.), by grants PCM-8316390 (to D.E.H.) and DMB-8316856 (to J.H.S.) from the National Science Foundation, by Public Health Service grants AI-10793 (to J.H.S.) and AI-20612 (to J.H.S.) from the National Institutes of Health, and by Biomedical Research Support grant 557 RR 07003. We thank Richard Maki for useful discussions and his generous gift of G418. Oregon State University Agricultural Experiment Station technical paper no. 7439.
Record Number:CaltechAUTHORS:FRAmcb85
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:FRAmcb85
Alternative URL:http://mcb.asm.org/cgi/content/abstract/5/8/1918
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ID Code:4866
Collection:CaltechAUTHORS
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Deposited On:11 Sep 2006
Last Modified:26 Dec 2012 09:02

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