Improved Transformation of the Filamentous Fungus Aspergillus niger Using Agrobacterium tumefaciens.
- Author:
Seung Moon PARK
1
Author Information
1. Basic Science Research Institute, Chonbuk National University, Chonju, Chonbuk 561-756, Korea. smpark@moak.chonbuk.ac.kr
- Publication Type:Original Article
- Keywords:
Aspergillus niger;
Fungal transformation
- MeSH:
Agar;
Agrobacterium tumefaciens*;
Agrobacterium*;
Aspergillus nidulans;
Aspergillus niger*;
Aspergillus*;
Fungi*;
Genome, Fungal;
Hygromycin B;
Niger;
Polymerase Chain Reaction;
Protoplasts;
Saccharomyces cerevisiae;
Spores, Fungal;
Yeasts
- From:Mycobiology
2001;29(3):132-134
- CountryRepublic of Korea
- Language:English
-
Abstract:
Since it is known that Agrobacterium tumefaciens, which has long been used to transform plants, can transfer the T-DNA to yeast Saccharomyces cerevisiae during tumourigenesis, a variety of fungi were subjected to transformation to improve their transformation frequency. In this study, I report the A. tumefaciens-mediated transformation of filamentous fungus Aspergillus niger. Transfer of the binary vector pBIN9-Hg, containing the bacterial hygromycin B phosphotransferase gene under the control of the Aspergillus nidulans trpC promoter and terminator as a selectable marker, led to the selection of 50~100 hygromycin B-resistant transformants per 1x10(7) conidia of A. niger. This efficiency is improved 10~20 fold more than reported elsewhere. In order to avoid the difficulties in selection transformant from the over-growing non-transformant, I used top agar containing 900 microg/ml of hygromycin. Genomic PCR and Southern analysis showed that all transformants contained single T-DNA insert per fungal genome. This technique offers an easier and more efficient method than that of using protoplast.
