Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 1990 Oct;64(10):4691-6.
doi: 10.1128/JVI.64.10.4691-4696.1990.

Linker insertion mutagenesis of herpesviruses: inactivation of single genes within the Us region of pseudorabies virus

N de Wind  1 , A Zijderveld, K Glazenburg, A Gielkens, A Berns
Affiliations

Linker insertion mutagenesis of herpesviruses: inactivation of single genes within the Us region of pseudorabies virus

N de Wind et al. J Virol. 1990 Oct.

Abstract

We describe a technique for the systematic inactivation of nonessential genes within the genome of a herpesvirus without the requirement for phenotypic selection. This technique is based on the insertion of an oligonucleotide containing translational stop codons at a random site within a large cloned viral DNA fragment. Mutant virus is then reconstituted by cotransfection with overlapping viral clones, together comprising the entire viral genome, as described previously (M. van Zijl, W. Quint, J. Briaire, T. de Rover, A. Gielkens, and A. Berns, J. Virol. 62:2191-2195, 1988). This technique was used to construct, in a single experiment, a set of 13 viable pseudorabies virus strains with oligonucleotide insertions within all known genes of the Us region except for the gp50 gene, which proved essential for virus growth in cell culture. The growth rate in porcine kidney cells of mutants of all nonessential Us genes was similar to that of the parental virus, with the exception of a mutant of the recently identified protein kinase gene.

PubMed Disclaimer

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Virology. 1975 Aug;66(2):385-92 - PubMed
    1. J Virol. 1988 Aug;62(8):2712-7 - PubMed
    1. Virology. 1979 Jun;95(2):285-94 - PubMed
    1. Methods Enzymol. 1980;65(1):415-26 - PubMed
    1. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2184-8 - PubMed

Publication types