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논문 상세정보

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

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.

Abstract

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.

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