A Rapid Method for Screening Vaccinia Virus Recombinants

Overview

Journal Biotechniques

Specialties Biomedical Engineering
Biotechnology

Date 1991 May 1

PMID 1680352

Citations 9

Authors

A C Lai

Y Chu

Affiliations

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Abstract

A rapid and small-scale method for screening vaccinia virus recombinants employing micrococcal nuclease is described. This protocol utilizes the differential sensitivity of cellular and viral DNA to the nuclease, which can be selectively activated by addition of Ca2+ and inactivated by elimination of Ca2+. Two to five micrograms of viral DNA can be obtained from one infected L cell plate (50 mm) after overnight incubation.

Citing Articles

Replication-inducible vaccinia virus vectors with enhanced safety in vivo.

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IL-18 expression results in a recombinant vaccinia virus that is highly attenuated and immunogenic.

Verardi P, Legrand F, Chan K, Peng Y, Jones L, Yilma T J Interferon Cytokine Res. 2013; 34(3):169-78.

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Modified H5 promoter improves stability of insert genes while maintaining immunogenicity during extended passage of genetically engineered MVA vaccines.

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Vaccinia viruses with a serpin gene deletion and expressing IFN-gamma induce potent immune responses without detectable replication in vivo.

Legrand F, Verardi P, Chan K, Peng Y, Jones L, Yilma T Proc Natl Acad Sci U S A. 2005; 102(8):2940-5.

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Induction of potent humoral and cell-mediated immune responses by attenuated vaccinia virus vectors with deleted serpin genes.

Legrand F, Verardi P, Jones L, Chan K, Peng Y, Yilma T J Virol. 2004; 78(6):2770-9.

PMID: 14990697 PMC: 353749. DOI: 10.1128/jvi.78.6.2770-2779.2004.