Identification and characterization of the Spodoptera littoralis nucleopolyhedrovirus type B lef-3 gene
| Autor: | D. B. Levin, José Luiz Caldas Wolff, L. M. Herzog, L. Sun |
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| Rok vydání: | 1998 |
| Předmět: |
DNA
Complementary animal structures Genes Viral Transcription Genetic viruses Molecular Sequence Data Gene Expression macromolecular substances Spodoptera Virus Replication Cell Line Viral Proteins Transcription (biology) Virology Gene expression Animals Amino Acid Sequence Northern blot Promoter Regions Genetic Gene Base Sequence biology fungi Chromosome Mapping General Medicine Oligonucleotides Antisense Blotting Northern biology.organism_classification Molecular biology Nucleopolyhedroviruses DNA-Binding Proteins Autographa californica Bromodeoxyuridine Viral replication DNA Viral embryonic structures Orgyia pseudotsugata Sequence Alignment |
| Zdroj: | Archives of Virology. 143:743-767 |
| ISSN: | 1432-8798 0304-8608 |
| DOI: | 10.1007/s007050050327 |
| Popis: | We have identified a gene from the Spodoptera littoralis nucleopolyhedrovirus type B (SpliNPV-B) with several characteristics that suggest that it is homologous to the lef-3 genes of the Autographa californica and Orgyia pseudotsugata NPVs (AcMNPV and OpMNPV, respectively). The SpliNPV-B lef-3 gene was mapped between 43.6 and 45.5 map units of the SpliNPV-B genome. Northern blot analysis showed that SpliNPV-B lef-3 was expressed as a 1.6 Kb transcript at 5 h post infection (p.i.), reached high levels at 24 h p.i., and remained highly expressed at 56 h p.i. Transcription of SpliNPV-B lef-3 initiated at two distinct sites downstream from a TATA-box motif and terminated 25 nucleotides downstream from the translation stop site of the putative LEF-3 polypeptide. The 5'-boundaries of lef-3 promoter elements were investigated by transient expression assays, which revealed that the major components of the lef-3 promoter are within a 183 base pair region upstream of the distal transcription initiation site. Transfection of SpliNPV-B infected Sf9 cells with anti-sense oligonucleotides designed to inhibit LEF-3 expression resulted in substantial reduction of viral DNA replication, suggesting that the role of SpliNPV-B lef-3 may be similar to that of AcMNPV and OpMNPV lef-3 genes, which are essential for viral DNA replication. |
| Databáze: | OpenAIRE |
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