title: Characterization of the ecdysteroid UDP-glucosyltransferase (egt) gene of Anticarsia gemmatalis nucleopolyhedrovirus.
abstract: The Anticarsia gemmatalis nucelopolyhedrovirus (AgMNPV) egt gene was cloned, sequenced and its expression characterized by RT-PCR and western blot analysis. Sequence analysis of the gene indicated the presence of an open reading frame (ORF) of 1482 nucleotides, which codes for a polypeptide of 494 amino acids. ATATA box and a conserved regulatory sequence (CATT) found in other baculovirus early genes were present in the promoter region of the egt gene. A poly-A consensus sequence was present in the 3' untranslated region (3'-UTR) of the gene. Homology comparisons showed that the EGT protein of AgMNPV is most closely related (95.9% amino acid sequence identity) to the EGT from the Choristoneura fumiferana DEF nucleopolyhedrovirus (CfDEF). Transcriptional analysis of the AgMNPV egt gene showed that egt-specific transcripts can be detected both early and late in infection. The EGT protein was detected, by western blot analysis, in the intra- (from 12 to 48 h post-infection) and extra-cellular (from 12 to 96 h post-infection) fractions of infected insect cells. The AgMNPV Bgl II-F fragment, which has homology to the AcMNPV ie-1 gene, was cloned and used to cotransfect SF21 cells with the cloned AgMNPV egt gene. EGT activity was observed, suggesting that AgMNPV ie-1 can transactivate egt expression.
authors.affiliation: Embrapa Genetic Resources and Biotechnology, Brasília, Brasil. carlyle@cenargen.embrapa.br
authors.forename: J C
authors.initials: JC
authors.lastname: Rodrigues
authors.forename: M L
authors.initials: ML
authors.lastname: De Souza
authors.forename: D
authors.initials: D
authors.lastname: O'Reilly
authors.forename: L M
authors.initials: LM
authors.lastname: Velloso
authors.forename: F J
authors.initials: FJ
authors.lastname: Pinedo
authors.forename: F B
authors.initials: FB
authors.lastname: Razuck
authors.forename: B
authors.initials: B
authors.lastname: Ribeiro
authors.forename: B M
authors.initials: BM
authors.lastname: Ribeiro
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002478:Cells, Cultured; D003001:Cloning, Molecular; D005964:Glucosyltransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D007915:Lepidoptera / Q000821:virology; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000201:enzymology* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D018411:Spodoptera / Q000821:virology; D014158:Transcription, Genetic
chemical_list: D005964:Glucosyltransferases; C086659:ecdysteroid UDP-glucosyltransferase