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Insect Mol Biol ; 12(5): 415-25, 2003 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-12974946

RESUMEN

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.


Asunto(s)
Anopheles/genética , Baculoviridae/genética , Escarabajos/genética , Expresión Génica , Spodoptera/enzimología , Transformación Genética , alfa-Amilasas/genética , Animales , Secuencia de Bases , Técnicas de Cultivo de Célula , Cromatografía , Cartilla de ADN , Electroforesis en Gel de Agar , Datos de Secuencia Molecular , Spodoptera/genética , alfa-Amilasas/metabolismo
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