Structure and properties of a herpesvirus of turkeys recombinant in which US1, US10 and SORF3 genes have been replaced by a lacZ expression cassette

In the process of generating an insertional mutant of herpesvirus of turkeys (HVT) expressing lacZ at the protein kinase (PK) locus, we isolated a recombinant which contained an intact PK gene but the short unique regions US1, US10 and SORF3 had been deleted and replaced by the lacZ cassette. Moreov...

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Bibliographic Details
Published in:Journal of general virology 1995-11, Vol.76 (11), p.2903-2907
Main Authors: Zelnik, V, Tyers, P, Smith, G.D, Jiang, C.L, Ross, N.L.J
Format: Article
Language:English
Subjects:
Animals
Antibodies, Viral - biosynthesis
Base Sequence
beta-galactosidase
Cells, Cultured
Chick Embryo
Chickens
Cloning, Molecular
deletions
DNA Primers
experimental infection
genbank/z30543
genbank/z30647
Gene Expression Regulation, Viral
Genes, Viral
genetic recombination
Herpesviridae - genetics
Herpesviridae - physiology
Lac Operon
lacz gene
Meleagrid herpesvirus 1
Molecular Sequence Data
Mutagenesis, Insertional
nucleotide sequences
Plasmids
recombinant DNA
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - immunology
Recombinant Fusion Proteins - metabolism
replication
reporter genes
structural genes
Turkeys - virology
Virus Replication
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Description
Summary:In the process of generating an insertional mutant of herpesvirus of turkeys (HVT) expressing lacZ at the protein kinase (PK) locus, we isolated a recombinant which contained an intact PK gene but the short unique regions US1, US10 and SORF3 had been deleted and replaced by the lacZ cassette. Moreover, the virus contained duplicate copies of gD, gI and gE in an opposite orientation flanking lacZ, US2 and PK which were contiguous. These results are of interest in relation to the flexibility of the short unique segment (Us) and of the inverted repeats flanking Us of the alpha-herpesviruses. The recombinant expressed beta-galactosidase and was genetically stable in vitro and in vivo. Chickens inoculated with the virus developed antibodies to HVT antigens and to beta-galactosidase but the replication of the recombinant in vivo was impaired in comparison to parental HVT as shown by a reduction in the proportion of infected lymphocytes.
ISSN:0022-1317
1465-2099
DOI:10.1099/0022-1317-76-11-2903