Development of an expression vector for Metarhizium anisopliae based on the tef-1alpha homologous promoter.
The high-conserved translation elongation factor 1 alpha (tef-1alpha) gene from the enthomopathogenic fungus Metarhizium anisopliae was characterized to select the promoter region. A 640-bp DNA fragment upstream to the start codon was employed to drive the expression of the reporter protein sGFP or a dominant selectable marker, the gene bar (resistance to ammonium glufosinate). Transformants carrying this homologous promoter system showed no difference in virulence bioassays against the cattle tick Boophilus microplus comparing to the M. anisopliae wild-type strain. Moreover, GFP fluorescence was detected during tick infection bioassay.
Nakazato
Luciano
L
Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, P.O. Box 15005, CEP 91501-970, Porto Alegre, RS, Brazil.
Dutra
Valéria
V
Broetto
Leonardo
L
Staats
Charley Christian
CC
Vainstein
Marilene Henning
MH
Schrank
Augusto
A
D000818:Animals; D001203:Ascomycota / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005260:Female; D005656:Fungal Proteins / Q000235:genetics; D005800:Genes, Fungal; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D026863:Ixodidae / Q000382:microbiology*; D008969:Molecular Sequence Data; D010445:Peptide Elongation Factors / Q000235:genetics*; D010802:Phylogeny; D011401:Promoter Regions, Genetic; D014170:Transformation, Genetic
D018076:DNA, Complementary; D005656:Fungal Proteins; D010445:Peptide Elongation Factors; D049452:Green Fluorescent Proteins