Nucleotide sequence of the Autographa californica nuclear polyhedrosis 9.4 kbp EcoR I-I and -R (Polyhedrin gene) region
Identifieur interne : 001E56 ( Istex/Checkpoint ); précédent : 001E55; suivant : 001E57Nucleotide sequence of the Autographa californica nuclear polyhedrosis 9.4 kbp EcoR I-I and -R (Polyhedrin gene) region
Auteurs : Robert D. Possee [Royaume-Uni] ; Tai-Ping Sun [Royaume-Uni] ; Stephen C. Howard [Royaume-Uni] ; Martin D. Ayres [Royaume-Uni] ; Michéle Hill-Perkins [Royaume-Uni] ; Katharine L. Gearing [Royaume-Uni]Source :
- Virology [ 0042-6822 ] ; 1991.
English descriptors
- Teeft :
- Academic press, Acmnpv, Acmnpv genome, Amino acids, Autographa, Autographa californica, Baculovirus, Baculovirus expression vectors, Baculovirus vector, Californica, Coding, Coding region, Coding sequence, Coding sequences, Codon, Complete sequence, Ecor, Endonuclease, Entire region, Foreign genes, Frugiperda, Frugiperda cells, Functional analysis, Gene, Gene encoding, Gene product, Genome, Hindill, Hindill restriction endonucleases, Hybridization, Hybridization analysis, Insect cells, Molecular biology, Mrna, Noncoding region, Northern blot hybridization analysis, Nucleic acids, Nucleotide, Nucleotide sequence, Oligonucleotide, Open reading frames, Orfs, Partial restriction, Plaque assay, Plaque purification, Plasmid, Polyhedrin, Polyhedrin coding sequences, Polyhedrin gene, Polyhedrosis, Polyhedrosis virus, Polyhedrosis virus genome, Possee, Present address, Primer, Primer extension analysis, Primer extension products, Progeny virus, Protein synthesis, Reading frames, Recombinant viruses, Restriction enzyme sites, Restriction enzymes, Rohrmann, Sequence analysis, Sequence data, Southern hybridization, Synthetic oligonucleotide, Temporal expression, Transcript, Transcription, Transcriptional analysis, Viral, Virology, Virus, Virus genome, Virus replication.
Abstract
Abstract: The nucleotide sequence of a 9.4-kbp region including the polyhedrin gene of the C6 strain of the Autographa californica nuclear polyhedrosis virus (AcMNPV) genome was determined. These data provide a complete description of the EcoRI-I fragment, which is used to produce transfer vectors for inserting foreign genes into the AcMNPV. Ten potential open reading frames (ORFs) were identified in the complete sequence, on either strand of DNA. The largest of these was 1629 nucleotides in length and was located downstream from the polyhedrin coding sequences, but on the opposite strand of DNA. Northern blot hybridization analysis of ORF 8 (1629) identified an RNA of 2000 nucleotides which was produced in infected cells from 12 hr p.i. and remained until at least 48 hr p.i. S1 nuclease mapping and analysis of cDNA clones located the 3′ end of the mRNA at a site 16 nucleotides downstream of the polyhedrin coding sequences. The 5′ end of the mRNA was mapped using primer extension analysis of polyadenylated RNA. ThemRNA start site was positioned within a late/very late consensus transcription initiation motif (ATAAG), 428 nucleotides upstream from the potential ATG translation initiation codon. The biological significance of the putative gene product was assessed by inserting a synthetic oligonucleotide in the carboxyl terminal coding sequences of ORF 8 (1629) to prematurely terminate translation. Recombinant viruses containing this mutation were not isolated, suggesting that the ORF 1629 gene product is essential for virus replication.
Url:
DOI: 10.1016/0042-6822(91)90770-C
Affiliations:
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These data provide a complete description of the EcoRI-I fragment, which is used to produce transfer vectors for inserting foreign genes into the AcMNPV. Ten potential open reading frames (ORFs) were identified in the complete sequence, on either strand of DNA. The largest of these was 1629 nucleotides in length and was located downstream from the polyhedrin coding sequences, but on the opposite strand of DNA. Northern blot hybridization analysis of ORF 8 (1629) identified an RNA of 2000 nucleotides which was produced in infected cells from 12 hr p.i. and remained until at least 48 hr p.i. S1 nuclease mapping and analysis of cDNA clones located the 3′ end of the mRNA at a site 16 nucleotides downstream of the polyhedrin coding sequences. The 5′ end of the mRNA was mapped using primer extension analysis of polyadenylated RNA. ThemRNA start site was positioned within a late/very late consensus transcription initiation motif (ATAAG), 428 nucleotides upstream from the potential ATG translation initiation codon. The biological significance of the putative gene product was assessed by inserting a synthetic oligonucleotide in the carboxyl terminal coding sequences of ORF 8 (1629) to prematurely terminate translation. Recombinant viruses containing this mutation were not isolated, suggesting that the ORF 1629 gene product is essential for virus replication.</div></front></TEI><affiliations><list><country><li>Royaume-Uni</li></country></list><tree><country name="Royaume-Uni"><noRegion><name sortKey="Possee, Robert D" sort="Possee, Robert D" uniqKey="Possee R" first="Robert D." last="Possee">Robert D. Possee</name></noRegion><name sortKey="Ayres, Martin D" sort="Ayres, Martin D" uniqKey="Ayres M" first="Martin D." last="Ayres">Martin D. Ayres</name><name sortKey="Gearing, Katharine L" sort="Gearing, Katharine L" uniqKey="Gearing K" first="Katharine L." last="Gearing">Katharine L. Gearing</name><name sortKey="Hill Perkins, Michele" sort="Hill Perkins, Michele" uniqKey="Hill Perkins M" first="Michéle" last="Hill-Perkins">Michéle Hill-Perkins</name><name sortKey="Howard, Stephen C" sort="Howard, Stephen C" uniqKey="Howard S" first="Stephen C." last="Howard">Stephen C. Howard</name><name sortKey="Sun, Tai Ping" sort="Sun, Tai Ping" uniqKey="Sun T" first="Tai-Ping" last="Sun">Tai-Ping Sun</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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