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Expression of functional Anopheles merus α‐amylase in the baculovirus/ Spodoptera frugiperda system
Author(s) -
Effio P. C.,
FolguerasFlatschart A. V.,
Montor W. R.,
Pernasetti F. M.,
Pueyo M. T.,
Sogayar M. C.
Publication year - 2003
Publication title -
insect molecular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.955
H-Index - 93
eISSN - 1365-2583
pISSN - 0962-1075
DOI - 10.1046/j.1365-2583.2003.00423.x
Subject(s) - biology , genbank , sf9 , spodoptera , accession number (library science) , gene , polyhedrin , signal peptide , microbiology and biotechnology , virology , genetics , recombinant dna
The Anopheles merus (Diptera, Nematocera, Culicoidea) α‐amylase gene (AmerAmy, GenBank Accession Number U01210 ) was amplified with its own or with the Zabrotes subfasciatus α‐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. subfasciatus α‐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™ system. Recombinant baculovirus preparations were used to infect Sf 9 Spodoptera frugiperda insect cells. The A. merus α‐amylase was successfully expressed as an active enzyme detected mainly in cell culture supernatants.