Gene inactivation mediated by Agrobacterium tumefaciens in the filamentous fungi Metarhizium anisopliae.
The list of fungal species with known complete genome and/or expressed sequence tag collections is extending rapidly during the last couple of years. Postgenomic gene function assignment is an obvious follow-up and depends on methodologies to test gene function in vivo. One of such methods is the generation of null mutants via homologous recombination at the wild-type loci by using inactivation cassettes. In this paper, the ability of Agrobacterium tumefaciens to genetically transform filamentous fungi was exploited to drive homologous recombination at the trp1 locus of the enthomopathogenic fungus Metarhizium anisopliae. The trp1 disruptants exhibited a clearly distinguishable phenotype from wild-type cells and were recovered with high efficiency of homologous recombination (22%). The complementation of such mutants with the wild-type gene generates only transformants with homologous integration.
Staats
Charley Christian
CC
Centro de Biotecnologia, Programa de Pós-graduação em Biologia celular e Molecular, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.
Junges
Angela
A
Fitarelli
Mariana
M
Furlaneto
Marcia Cristina
MC
Vainstein
Marilene Henning
MH
Schrank
Augusto
A
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D019747:Aldose-Ketose Isomerases / Q000235:genetics*; D018920:Coculture Techniques; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics*; D020868:Gene Silencing; D052981:Metarhizium / Q000235:genetics*; D008855:Microscopy, Electron, Scanning; D016254:Mutagenesis, Insertional; D010641:Phenotype; D011995:Recombination, Genetic
D004269:DNA, Bacterial; D005656:Fungal Proteins; C036483:T-DNA; D019747:Aldose-Ketose Isomerases