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Chemically-induced temperature sensitive mutants of dengue virus type 2

I. Isolation and partial characterization

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Summary

Temperature sensitive(ts) mutants of dengue virus type 2 (DEN-2, TH-36 isolate) were induced by replication in primary hamster kidney cells treated with 5-azacytidine. Sevents mutants were obtained from 138 clones isolated by an immunofluorescent cloning technique. Of these 7ts mutants, 5 were sufficiently stable to permit partial characterization. Complementation was detected at very low but statistically significant levels between somets mutants at 40° C. Viral double-stranded RNA production was evaluated in LLC-MK2 cells at 30° and 40° C by micro-quantitative complement fixation. The results of complementation tests and RNA production tests indicated that the 4 of 5 stablets mutants constitute 3 separate complementation groups (2 RNA+ and 1 RNA− groups), while a fifthts mutant was RNA− but non-complementable. The data presented here indicate that a genetic system can be developed without employing traditional plaque or cytopathology methods. Further, the 5 DEN-2ts mutants are believed to represent the only set of complementation-positive flavivirus mutants so far isolated.

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Authors and Affiliations

  1. Department of Microbiology, Graduate School of Public Health, University of Pittsburgh, Pittsburgh, Pennsylvania, USA

    G. C. Tarr & A. S. Lubiniecki

  2. Life Sciences Division Meloy Laboratories, Inc., Springfield, Virginia, USA

    G. C. Tarr & A. S. Lubiniecki

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  1. G. C. Tarr
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  2. A. S. Lubiniecki
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With 1 Figure

This research was supported, in part, by the U.S. Army Medical Research and Development Command under Contract Nos. DADA-16-69-C-9048 and DAMD-17-74-C-4005, and by the National Institute of Allergy and Infectious Diseases, National Institutes of Health, U.S. Public Health Service, under Research Grant Nos. 5-R22-AI02686-13,-14, and -15, and Training Grant No. 5-T01-AI-00110-14.

This work was begun while at the University of Pittsburgh and continued to a major extent at the present address.

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Tarr, G.C., Lubiniecki, A.S. Chemically-induced temperature sensitive mutants of dengue virus type 2. Archives of Virology 50, 223–235 (1976). https://doi.org/10.1007/BF01320576

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