Improvement on genetic transformation in the nematode-trapping fungus Arthrobotrys oligospora and its quantification on dung samples.

Abstract:

:An improved DNA-mediated transformation system for nematode-trapping fungus Arthrobotrys oligospora based on hygromycin B resistance was developed. The transformation frequency varied between 34 and 175 transformants per microg linearized DNA and 93% of the transformants were stable for drug resistance when tested 100 randomly selected transformants. More than 2000 transformants were obtained by transformation of the fungus with pBChygro in the presence of HindIII and among them, one, YMF1.00110, which lost its ability of forming predacious structure, was isolated. Southern analysis showed that the plasmid DNA had integrated into the genome of all tested transformants (including YMF 1.00110) except one. The transformant tagged with hph gene could be re-isolated and quantified from dung samples based on the resistance of hygromycin B. All the results suggested that the method of restriction enzyme mediated integration (REMI) should facilitate not only the insertional mutagenesis for tagging and analysis genes of interest but also the ecological investigation of tagged fungi in a given environment.

journal_name

Mycopathologia

journal_title

Mycopathologia

authors

Jin X,Ming-He M,Xiao-Wei H,Ke-Qin Z

doi

10.1007/s11046-005-4334-2

keywords:

subject

Has Abstract

pub_date

2005-06-01 00:00:00

pages

533-8

issue

4

eissn

0301-486X

issn

1573-0832

journal_volume

159

pub_type

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