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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    (전북대학교 생물과학부 및 유전공학연구소  );
  • 초록

    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.


    A mutant (HSMl) of Cryphonectria parasitica created during transformation reproduced the hypovirulent symptoms in virus-free wild type. Its phenomena have been proved with morphological marker such as reduced sporulation, pigmentation, and laccase production. In addition to the changes in phenotypic characteristics, down-regulations of Lac1, Crp1, Vir1 and Vir2 were also observed. The integration of transforming vector was confirmed and located within genome by marker rescuing. Vector integration occurred between two genes, Cpg2 and Cpg3, which resulted in the disruption of neither Cpg2 nor Cpg3. Both Cpg2 and Cpg3 genes, sized at 1.8 kb and 1.9 kb respectively, were rarely transcribed genes in Cryphonectria parasitica. Cpg2 expression was significantly overexpressed from 4 to 5 day old culture of both UEP1 and HSM1 while no differences were observed in Cpg3 expression. It appears that an aberration from the normal expression of Cpg2, not Cpg3, results in the hypovirulent symptoms in virus-free wild type.


  • 주제어

    Cryphonectria parasitica .   Hypovirus .   Marker rescuing.  

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 저자의 다른 논문

  • Kim, Dae-Hyuk (33)

    1. 1997 "Induced Change in DNA Methylation of Fusarium oxysporum f. sp. niveum due to Successive Transfer" Journal of biochemistry and molecular biology = 한국생화학회지 30 (3): 216~221    
    2. 1998 "Cryparin 유전자의 promoter 분석을 위한 cryparin 유전자 치환체의 순수 제조" 한국균학회지 = The Korean journal of mycology 26 (4): 450~457    
    3. 1998 "Analysis of Genetic Variation in Botrytis cinerea Isolates Using Random Amplified Polymorphic DNA Markers" Journal of microbiology and biotechnology 8 (5): 490~496    
    4. 2000 "Expression of Murine GM-CSF in Recombinant Aspergillus niger" Journal of microbiology and biotechnology 10 (3): 287~292    
    5. 2000 "Ordered Differential Display from Cryphonectria parasitica" The plant pathology journal 16 (3): 142~146    
    6. 2001 "Expression of Nutritionally Well-balanced Protein, AmA1, in Saccharomyces cerevisiae" Biotechnology and bioprocess engineering 6 (3): 173~178    
    7. 2002 "Characterization of Aspergillus niger Mutants Deficient of a Protease" Mycobiology 30 (3): 160~165    
    8. 2003 "Modulation of a Fungal Signaling by Hypovirus" The plant pathology journal 19 (1): 30~33    
    9. 2003 "Production of a Functional Mouse Interferon ${\gamma}$from Recombinant Saccharomyces cerevisiae" Journal of microbiology and biotechnology 13 (4): 537~543    
    10. 2003 "Ferritin 유전자 전이 효모(Saccharomyces serevisiae)의 급여가 닭의 생산성, 장기 및 계란의 철분함량에 미치는 영향" 한국가금학회지 = Korean journal of poultry science 30 (4): 245~251    

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