Herpes simplex virus recombination vectors designed to allow insertion of modified promoters into transcriptionally "neutral" segments of the viral genome.
The use of recombinant viruses has been essential in investigation of the biology of herpes simplex virus (HSV). In this communication we describe a number of viral recombination vectors that we have generated for use in promoter structure/function analysis within the context of the HSV-1 genome. We have utilized two regions of the HSV genome that contain genes nonessential for replication in cultured cells--the glycoprotein C (gC or UL44) locus in the UL of the genome and the area encompassing the promoter and 5' portion of the latency associated transcript (LAT) within the RL factual influence on promoters due to the site of insertion. Two different kinetic promoters were analyzed, those controlling expression of the gamma UL 38 and the beta dUTPase genes, in both loci. All constructs tested displayed reporter gene mRNA expression with expected kinetics, and we conclude that there are no neighboring cryptic promoter elements that could interfere with expression studies using the vectors described.
Singh
J
J
Department of Molecular Biology and Biochemistry, University of California, Irvine 92717, USA.
Wagner
E K
EK
D000818:Animals; D001483:Base Sequence; D002522:Chlorocebus aethiops; D004274:DNA, Recombinant; D004279:DNA, Viral; D017930:Genes, Reporter; D005822:Genetic Vectors; D016679:Genome, Viral; D018259:Herpesvirus 1, Human / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D011401:Promoter Regions, Genetic; D011755:Pyrophosphatases / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D014158:Transcription, Genetic; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics
D004274:DNA, Recombinant; D004279:DNA, Viral; D012333:RNA, Messenger; D014759:Viral Envelope Proteins; C038535:glycoprotein gC, herpes simplex virus type 1; D001616:beta-Galactosidase; D011755:Pyrophosphatases; C023278:uridine triphosphatase