MUTAGENESIS AND CHARACTERIZATION OF A 41-KILOBASE-PAIR REGION OF THE PSEUDORABIES VIRUS GENOME - TRANSCRIPTION MAP, SEARCH FOR VIRULENCE GENES, AND COMPARISON WITH HOMOLOGS OF HERPES-SIMPLEX VIRUS TYPE-1

被引:24
作者
DEWIND, N
PEETERS, BPH
ZIJDERVELD, A
GIELKENS, ALJ
BERNS, AJM
KIMMAN, TG
机构
[1] CENT VET INST,DEPT VIROL,LELYSTAD,NETHERLANDS
[2] NETHERLANDS CANC INST,1066 CX AMSTERDAM,NETHERLANDS
关键词
D O I
10.1006/viro.1994.1242
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
We mutagenized and characterized a 41-kilobase-pair subgenomic cloned fragment from the unique long (UL) region of pseudorabies virus (PRV). Forty mutant clones, each carrying a single inserted oligonucleotide, were used for cotransfection with overlapping cloned viral DNA fragments. More than half of these transfections yielded viable virus mutants. Short viral DNA fragments, flanking the oligonucleotide insertions, were cloned and used as probes on Northern blots with RNA isolated from cells infected with wild-type PRV. In this way we were able to construct a partial transcriptional map of this region of the PRV genome. In addition, we used these probes in cross-hybridization studies with cloned genomic fragments from the prototype alphaherpesvirus herpes simplex virus type 1 (HSV-1). This allowed us to define homology between the corresponding regions of these viruses. Most viable PRV mutants were assayed for virulence in mice. Mutagenesis of the identified homologs of HSV-1 genes UL39 (encoding the large subunit of ribonucleotide reductase), UL40 (small subunit of ribonucleotide reductase), UL42 (DNA polymerase accessory factor), UL23 (thymidine kinase), UL21 (a capsid-associated protein), and UL12 (alkaline nuclease) completely abrogated or strongly reduced virulence. (C) 1994 Academic Press, Inc.
引用
收藏
页码:784 / 790
页数:7
相关论文
共 38 条
[1]  
Baskerville A., 1973, Veterinary Bulletin, V43, P465
[2]   LOCALIZATION OF THE REGIONS OF HOMOLOGY BETWEEN THE GENOMES OF HERPES-SIMPLEX VIRUS, TYPE-1, AND PSEUDORABIES VIRUS [J].
BENPORAT, T ;
VEACH, RA ;
IHARA, S .
VIROLOGY, 1983, 127 (01) :194-204
[3]   PROTEINS SPECIFIED BY THE SHORT UNIQUE REGION OF THE GENOME OF PSEUDORABIES VIRUS PLAY A ROLE IN THE RELEASE OF VIRIONS FROM CERTAIN CELLS [J].
BENPORAT, T ;
DEMARCHI, J ;
PENDRYS, J ;
VEACH, RA ;
KAPLAN, AS .
JOURNAL OF VIROLOGY, 1986, 57 (01) :191-196
[4]  
BENPORAT T, 1985, HERPESVIRUSES, V3, P105
[5]   THE PSEUDORABIES VIRUS HOST-SHUTOFF HOMOLOG GENE - NUCLEOTIDE-SEQUENCE AND COMPARISON WITH ALPHAHERPESVIRUS PROTEIN COUNTERPARTS [J].
BERTHOMME, H ;
JACQUEMONT, B ;
EPSTEIN, A .
VIROLOGY, 1993, 193 (02) :1028-1032
[6]   AN UNUSUAL SPLICED HERPES-SIMPLEX VIRUS TYPE-1 TRANSCRIPT WITH SEQUENCE HOMOLOGY TO EPSTEIN-BARR VIRUS-DNA [J].
COSTA, RH ;
DRAPER, KG ;
KELLY, TJ ;
WAGNER, EK .
JOURNAL OF VIROLOGY, 1985, 54 (02) :317-328
[7]   LOCATION AND ORIENTATION OF HOMOLOGOUS SEQUENCES IN THE GENOMES OF 5 HERPESVIRUSES [J].
DAVISON, AJ ;
WILKIE, NM .
JOURNAL OF GENERAL VIROLOGY, 1983, 64 (SEP) :1927-1942
[8]   HERPESVIRUSES ENCODE AN UNUSUAL PROTEIN-SERINE THREONINE KINASE WHICH IS NONESSENTIAL FOR GROWTH IN CULTURED-CELLS [J].
DEWIND, N ;
DOMEN, J ;
BERNS, A .
JOURNAL OF VIROLOGY, 1992, 66 (09) :5200-5209
[9]   RIBONUCLEOTIDE REDUCTASE-DEFICIENT MUTANTS OF PSEUDORABIES VIRUS ARE AVIRULENT FOR PIGS AND INDUCE PARTIAL PROTECTIVE IMMUNITY [J].
DEWIND, N ;
BERNS, A ;
GIELKENS, A ;
KIMMAN, T .
JOURNAL OF GENERAL VIROLOGY, 1993, 74 :351-359
[10]   THE PSEUDORABIES VIRUS HOMOLOG OF THE HERPES-SIMPLEX VIRUS UL21 GENE-PRODUCT IS A CAPSID PROTEIN WHICH IS INVOLVED IN CAPSID MATURATION [J].
DEWIND, N ;
WAGENAAR, F ;
POL, J ;
KIMMAN, T ;
BERNS, A .
JOURNAL OF VIROLOGY, 1992, 66 (12) :7096-7103