Transformation of the nematode-trapping fungus Arthrobotrys oligospora

被引:62
作者
Tunlid, A
Åhman, J
Oliver, RP
机构
[1] Lund Univ, Dept Microbial Ecol, S-22362 Lund, Sweden
[2] Carlsberg Lab, Dept Physiol, DK-2500 Copenhagen, Denmark
关键词
nematode-trapping fungi; transformation; hygromycin resistance; restriction mediated integration; beta-glucuronidase reporter gene;
D O I
10.1111/j.1574-6968.1999.tb13491.x
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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/CaCl2 treatment. Transformation frequencies varied between 1-6 transformants per mu g 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 uidA gene encoding the beta-glucuronidase reporter gene under the control of an Aspergillus nidulans promoter, occurred at frequencies of up to 63%. (C) 1999 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.
引用
收藏
页码:111 / 116
页数:6
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