High efficient homologous integration via tandem exo-beta-1,3-glucanase genes in the common mushroom, Agaricus bisporus

Homologous integration was studied in the common mushroom, Agaricus bisporus, using a plasmid (pHAG3-1) carrying the hygromycin-resistance gene and a 3.2-kb genomic fragment from A. bisporus. Homologous integration was found in 30-60% of the transformants obtained with pHAG3-1 linearized at three di...

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Bibliographic Details
Published in:Current genetics 1996, Vol.30 (2), p.166-173
Main Authors: Rhee, M.D. van de, Mendes, O, Werten, M.W.T, Huizing, H.J, Mooibroek, H
Format: Article
Language:English
Subjects:
abexg1 gene
abexg2 gene
Agaricus bisporus
amino acid sequences
codon usage
fruiting bodies
fungal anatomy
genbank/x92961
gene expression
genetic code
genetic transformation
homologous recombination
introns
linearized plasmids
messenger RNA
molecular sequence data
mycelium
nucleotide sequences
O-glycoside hydrolases
open reading frames
plasmid vectors
restriction mapping
structural genes
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Summary:Homologous integration was studied in the common mushroom, Agaricus bisporus, using a plasmid (pHAG3-1) carrying the hygromycin-resistance gene and a 3.2-kb genomic fragment from A. bisporus. Homologous integration was found in 30-60% of the transformants obtained with pHAG3-1 linearized at three different positions within the homologous sequence, generating either blunt, 5'- or 3'-protruding ends. The genomic fragment was found to contain two homologous open reading frames in tandem, which showed 60% similarity to exo-beta-1,3-glucanases from Saccharomyces cerevisiae and Candida albicans. The level of the corresponding mRNA is low in the vegetative mycelium and relatively high in fruiting bodies. In the vegetative mycelium of a transformant with tandemly integrated pHAG3-1 plasmids at the homologous position, exoglucanase mRNA was strongly increased without any apparent effect on growth rate or morphology.
ISSN:0172-8083
1432-0983