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Publications (4)6.2 Total impact

  • P C Hartig, M C Cardon
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    ABSTRACT: Recombinant baculoviruses have been used to produce foreign proteins and have the potential to be safe, efficacious insecticides. Isolation of recombinant virus is usually by plaque phenotype. Typical recombination rates are less than 1%, thus requiring time consuming inspection of hundreds of individual plaques. We describe a method of generating recombinants which requires less time than current protocols and frequently produces recombinants at rates exceeding 30%. This protocol employs liposome-mediated transfection, reduced post-transfection incubation times, linearized parental virus which produces occlusion positive plaques in clones of the parental genotype, and colorimetric detection of recombinants. This protocol allows the initial, and frequently the final, isolation of recombinants in 7 days.
    Journal of Virological Methods 08/1992; 38(1):61-70. · 1.90 Impact Factor
  • P C Hartig, M C Cardon, C Y Kawanishi
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    ABSTRACT: Risk assessment of viral pesticides is required by federal statute. Subdivision M of the Pesticide Testing Guidelines includes mammalian cell culture studies for viral agents. Using these tests we have demonstrated baculovirus toxicity to human fibroblast and lung cells, as well as monkey kidney cells. However, virus was not toxic to toad cells. Furthermore, toxicity was associated with the virus particle and was not removed by dialysis of the preparation, gradient purification or psoralen inactivation of the virus. However, heat-inactivated virus was not toxic.
    Developments in biological standardization 02/1992; 76:313-7.
  • P C Hartig, M C Cardon, C Y Kawanishi
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    ABSTRACT: Baculovirus expression vectors have become a popular method of producing recombinant proteins. Production of recombinant virus requires the transfection of both the native viral DNA and a transfer plasmid into insect cells where recombination takes place. While several methods of transfecting insect cells exist, we have found liposome-mediated transfection to be highly efficient. Here we detail the protocols and medium needed for efficient, simple transfection of Spodoptera frugiperda cells.
    BioTechniques 10/1991; 11(3):310, 312-3. · 2.40 Impact Factor
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    ABSTRACT: Viral pesticidal agents must be evaluated for potential health hazards prior to utilization. Assessment of the likelihood of replication in humans has included in vitro exposure of human cells to the potential pesticidal agent. Previous in vitro evaluation strategies have lacked positive controls. Thus, negative results, interpreted as no effect of the virus on human cells, could reflect basic deficiencies in the testing protocols. We designed a testing scheme for viral pesticides and used it to test the nuclear polyhedrosis virus of Autographa californica. Tests were aimed at evaluating potential replication or gene expression in primate cells. Parallel tests were run utilizing identical protocols with primate viruses known to produce the biological effect being evaluated. Thus protocols described were tested with positive viral controls.
    Journal of Virological Methods 02/1991; 31(2-3):335-44. · 1.90 Impact Factor