Open AccessTransformation of the nematode‐trapping fungus Arthrobotrys oligospora
Author(s) -
Tunlid A,
Åhman J,
Oliver R.P
Publication year - 1999
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1999.tb13491.x
Subject(s) - transformation (genetics) , aspergillus nidulans , biology , hygromycin b , plasmid , microbiology and biotechnology , protoplast , gene , escherichia coli , dna , promoter , reporter gene , genetics , gene expression , mutant
The nematode‐trapping fungus Arthrobotrys oligospora was transformed to hygromycin resistance using the hygromycin‐B phosphotransferase gene from Escherichia coli under the control of various heterologous fungal promoters. Plasmid DNA was introduced into fungal protoplasts by polyethylene glycol/CaCl 2 treatment. Transformation frequencies varied between 1–6 transformants per μg DNA. Seven out of 13 integration events analyzed from transformants were single copy integrations, whereas the remaining were multiple and more complex integrations. The addition of restriction enzymes during transformations increased the frequency of single copy integrations. Co‐transformation, using the E. coli uid A gene encoding the β‐glucuronidase reporter gene under the control of an Aspergillus nidulans promoter, occurred at frequencies of up to 63%.