Open AccessTransformation of the fungus Absidia glauca by complementation of a methionine‐auxotrophic strain affected in the homoserine‐acetyltransferase gene
Author(s) -
Karimi Sedighe,
Wetzel Jana,
Wöstemeyer Johannes,
Burmester Anke
Publication year - 2012
Publication title -
febs open bio
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.718
H-Index - 31
ISSN - 2211-5463
DOI - 10.1016/j.fob.2012.07.002
Subject(s) - complementation , auxotrophy , plasmid , transformation (genetics) , biology , mutant , homoserine , gene , protein fragment complementation assay , genetics , methionine , acetyltransferase , virulence , quorum sensing , acetylation , amino acid
Transformation of fungi by complementation of auxotrophs is generally much more reliable than usage of antibiotic resistance markers. In order to establish such a system for the model zygomycete Absidia glauca , a stable methionine auxotrophic mutant was isolated after X‐ray mutagenesis of the minus mating type and characterized at the molecular level. The mutant is disrupted in the coding region of the Met2‐1 gene, encoding homoserine O‐acetyltransferase. The corresponding wild type gene was cloned, sequenced and inserted into appropriate vector plasmids. Transformants are prototrophs and show restored methionine‐independent growth, based on complementation by the autonomously replicating plasmids.