RESUMO
To improve the expression level of heterologous genes in Flammulina velutipes Fv-1, we constructed new vectors having glyceraldehydes-3-phosphate dehydrogenase (gpd) gene promoter to control the expression of target genes. When the hygromycin B phosphotransferase (hph) gene from Escherichia coli was controlled by the gpd promoter, transformation efficiency was 3-fold higher than the case of that controlled by the tryptophan synthetase gene (trp1) promoter.
Assuntos
Flammulina/genética , Engenharia Genética/métodos , Gliceraldeído-3-Fosfato Desidrogenases/genética , Regiões Promotoras Genéticas/genética , Transformação Genética/genética , Escherichia coli/enzimologia , Escherichia coli/genética , Fosfotransferases (Aceptor do Grupo Álcool)/genética , Plasmídeos/genéticaRESUMO
To develop a gene transformation method for Flammulina velutipes, we constructed a vector with hph gene under control of the trp1 gene promoter. The vector was integrated into protoplast derived from mycelia by the calcium-polyethylene glycol method, as it has not been reported for F. velutipes. Transformation efficiency was much improved when transformation was performed by the restriction enzyme mediated integration method.
