|
KMID : 0364820140500040372
|
|
Korean Journal of Microbiology
2014 Volume.50 No. 4 p.372 ~ p.375
|
|
|
Protoplast Isolation and Genetic Transformation of Polyporus brumalis
|
|
You Seon-Hwa
Kim Myung-Kil
|
|
|
Abstract
|
|
|
This experiment was undertaken to investigate proper conditions for protoplast isolation and genetic transformation
of the white rot fungi, Polyporus brumalis. The protoplasts were formed from mycelia at a frequency of 1 × 107
/ml with 0.5% Usukizyme. The transformation vector (pHYgpt) was constructed using hygromycin resistance gene (hph)
for the selectable maker. The yield was 100?160 transformants/μg DNA in a transformation mediated by 40%
polyethylene glycol solution with aurintricarboxylic acid, heparin and supermidine. The genomic integration of the
pHYgpt was confirmed by hph-specific PCR and the expected amplified band appeared only in the transformants.
These results could be an efficient tool in gene engineering of the genus polyporus.
|
|
|
KEYWORD
|
|
|
Polyporus brumalis, protoplast, pYHgpt, transformation, white rot fungi
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|