|
KMID : 1160220010290030132
|
|
Mycobiology
2001 Volume.29 No. 3 p.132 ~ p.134
|
|
|
Improved Transformation of the Filamentous Fungus Aspergillus niger Using Agrobacterium tumefaciens
|
|
Park Seung-Moon
|
|
|
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 μg/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.
|
|
|
KEYWORD
|
|
|
Aspergillus niger, Fungal transformation
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
 
|