Earticle

초록

영어

Ceriporiopsis subvermispora is a unique white rot fungus degrading plant cell wall lignin without severe loss of cellulose. Recombinant plasmids containing homologous gene expression signals fused to the coding sequence of Escherichia coli hph which encodes for hygromycin phosphotransferase were introduced in protoplasts of wild type C. subvermispora using PEG/CaCl2 protocol. A number of hygromycin B resistant strains were isolated on a screening plate containing 100㎍/ml of hygromycin B: whereas, no colonies were observed when protoplasts were treated with no DNA as a negative control. It was demonstrated that most of the isolates lost their drug resistance during successive cultivations in the presence of the same concentration of hygromycin B, but some of the isolates showed stable drug resistance after five times repeated screening. They did not lose the drug resistance even after the cultivation in the absence of hygromycin B and incorporation of the hph sequence was confirmed by specific amplification of the target sequence in PCR experiments and Southern hybridization analysis. The stable transformation system will make it possible to do molecular genetic analysis, as well as breeding of genetically modified strains, in C. subvermispora. Moreover, it was demonstrated that recombinant constructs with truncated promoter showed reduced number of the drug resistant isolates on the first screening plate, in response with the length of the remaining promoter sequence. These findings indicated that unstable drug resistance observed in these isolates should originate from transient expression of the introducing marker genes, and that a promoter assay system has been developed for the first time in basidiomycete. This system is practically not affected by the positional effect of the integrated recombinant gene in the host chromosome.

저자

• Yoichi Honda [ RISH, Kyoto University, Gokasho, Uji, Japan ]
• Eiji Tanigawa [ RISH, Kyoto University, Gokasho, Uji, Japan ]
• Takahito Watanabe [ RISH, Kyoto University, Gokasho, Uji, Japan ]
• Takashi Watanabe [ RISH, Kyoto University, Gokasho, Uji, Japan ]

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  한국버섯학회지 [Journal of MUSHROOMS]

• 간기

  계간

• pISSN

  1738-0294

• eISSN

  2288-8853

• 수록기간

  2003~2025

• 등재여부

  KCI 등재

• 십진분류

  KDC 525 DDC 635