Identification, sequence, and transcriptional analysis of lef-3, a gene essential for Orgyia pseudotsugata baculovirus DNA replication.
Using a transient complementation assay for DNA replication of an origin-containing reporter plasmid, a 2.4-kb region (m.u. 48.3-50.1) of the Orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (OpMNPV) genome was identified that contains a gene essential for OpMNPV DNA replication. This region was sequenced and open reading frames were identified. Replication assays using subclones from this region identified a gene called late expression factor 3 (lef-3), as the essential replication gene. Transcriptional mapping using both Northern blot and S1 nuclease protection assays demonstrated that lef-3 was expressed as an early gene with a major transcript of 1.7 kb in length. OpMNPV lef-3 encodes a protein with a predicted molecular weight of 42.6 kDa (373 amino acids) and exhibits 41% amino acid sequence identity with its homolog in the genome of the Autographa californica MNPV. A pattern of amino acids similar to that found in single-stranded DNA binding proteins is present in the amino-terminus of both OpMNPV and AcMNPV LEF-3.
Ahrens
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Genetics Program, Oregon State University, Corvallis 97331-7301, USA.
Carlson
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Rohrmann
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D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004261:DNA Replication / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics*; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral / Q000235:genetics*; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000235:genetics* / Q000502:physiology; D016366:Open Reading Frames / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D012367:RNA, Viral / Q000032:analysis; D018741:Replication Origin / Q000235:genetics; D015183:Restriction Mapping; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014764:Viral Proteins / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics
D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D012367:RNA, Viral; D014764:Viral Proteins; C094647:late expression factor 3, baculovirus