• 제목/요약/키워드: Marker rescuing

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Cryphonectria parasitica에서 mycovirus 감염 증상을 보이는 돌연변이 균주의 선발과 marker rescuing을 이용한 돌연변이의 특성화 (Selection and Characterization of the Hypovirulent Symptom Micmicking Mutant in Cryphonectria parasitica Using Marker Rescuing)

  • 김대혁
    • 한국균학회지
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    • 제25권3호통권82호
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    • pp.191-201
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    • 1997
  • 유전자의 삽입에 의해 발생하는 C. parasitica의 돌연변이체중 mycovirus에 감염된 것과 같이 색소와 포자를 적게 형성하는 균주(HSM1)를 선발하였다. 선발된 균주는 형태학적 병징외에도 laccase효소의 역가와 같은 생화학적 그리고 표지 유전자들을 통해 분자 생물학적인 특징도 virus에 감염된 균주와 동일한 특징을 나타냈다. HSM1에서 돌연변이가 일어날 부위를 cloning하여 조사한 결과, 유전자 삽입 부위는 C. parasitica의 두 유전자(Cpg2와Cpg3)의 사이(intergenic space)이며 유전자의 삽입 결과, HSM1에서 Cpg2의 발현이 오히려 증가됨이 관찰되었고, 나아가 이와 같은 현상은 mycovirus 감염 균주(UEP1)에서도 일어나고 있음을 확인하였다.

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Protoplast-Mediated Transformation of the Filamentous Fungus Cladosporium phlei: Evidence of Tandem Repeats of the Integrative Transforming Vector

  • Kim, Jung-Ae;Kim, Jung-Mi;Kim, Hwan-Gyu;Kim, Beom-Tae;Hwang, Ki-Jun;Park, Seung-Moon;Yang, Moon-Sik;Kim, Dae-Hyuk
    • The Plant Pathology Journal
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    • 제25권2호
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    • pp.179-183
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    • 2009
  • To facilitate the genetic manipulation of Cladosporium phlei, a causal agent of leaf spot disease in timothy (Phleum pretense), protoplast-mediated transformation of C. phlei has been developed and the resulting transformants were characterized in this study. Hygromycin B resistance was applied as a dominant selection marker due to the sensitivity of C. phlei to this antibiotic. The transformation efficiency ranged from approximately 20-100 transformants per experiment. Southern blot analysis of stable transformants revealed that transformation occurred by way of stable integration of the vector DNA into the fungal chromosome. PCR analysis and plasmid rescuing of randomly selected transformants suggested that integration of tandem repeat copies of vector DNA was common. In addition, multiple integrations of the transforming vector at different chromosomal sites were also observed. The establishment of a transformation method for C. phlei facilitates strain improvement of this fungus and can be applied as an initial step in the molecular analysis of pigment production in this fungus.