title: Expression of functional Anopheles merus alpha-amylase in the baculovirus/Spodoptera frugiperda system.
abstract: The Anopheles merus (Diptera, Nematocera, Culicoidea) alpha-amylase gene (AmerAmy, GenBank Accession Number U01210) was amplified with its own or with the Zabrotes subfasciatusalpha-amylase signal peptide (ZsAmerAmy, GenBank Accession Number AY270183) by PCR, using designed primers. The AmerAmy gene was sequenced from its promotor to the TGA codon. As a positive control, the Z. subfasciatusalpha-amylase gene with its own signal peptide (ZsAmy, GenBank Accession Number AF255722) was also amplified by PCR. These three sequences were inserted into the baculovirus genome using the Bac-to-Bac trade mark system. Recombinant baculovirus preparations were used to infect Sf9 Spodoptera frugiperda insect cells. The A. merusalpha-amylase was successfully expressed as an active enzyme detected mainly in cell culture supernatants.
authors.affiliation: Instituto de Quimica, Universidade de São Paulo, São Paulo, Brazil.
authors.forename: P
authors.initials: P
authors.lastname: Chimoy Effio
authors.forename: A V
authors.initials: AV
authors.lastname: Folgueras-Flatschart
authors.forename: W R
authors.initials: WR
authors.lastname: Montor
authors.forename: F M
authors.initials: FM
authors.lastname: Pernasetti
authors.forename: M T
authors.initials: MT
authors.lastname: Pueyo
authors.forename: M C
authors.initials: MC
authors.lastname: Sogayar
authors.forename: P C
authors.initials: PC
authors.lastname: Effo
mesh_terms: D000818:Animals; D000852:Anopheles / Q000235:genetics*; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D018929:Cell Culture Techniques; D002845:Chromatography; D001517:Coleoptera / Q000235:genetics*; D017931:DNA Primers; D004587:Electrophoresis, Agar Gel; D015870:Gene Expression; D008969:Molecular Sequence Data; D018411:Spodoptera / Q000201:enzymology* / Q000235:genetics; D014170:Transformation, Genetic; D000516:alpha-Amylases / Q000235:genetics* / Q000378:metabolism
chemical_list: D017931:DNA Primers; D000516:alpha-Amylases