Trichoderma harzianum의 형질전환을 위한 Agrobacterium의 이용
Use of Agrobacterium for the Genetic Transformation of Trichoderma harzianum
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초록
Trichoderma harzianum 균사체를 Al_2O_3 입자와 마찰시킴으로써 Agrobacterium을 이용한 형질전환에 효과적으로 이용할 수 있었다. Hygromycin 저항성 균사체 출현을 비교한 결과 형질전환 효율이 20% 정도로 나타났으며 대조군의 경우 형질전환균사체의 출현은 없었다. 2차례의 연속적인 항생제배지에서 선발을 거친 형질전환균사체들은 PCR에 의하여 안정적 DNA도입이 확인되었으며 RT-PCR에 의하여 target gene의 mRNA발현을 확인할 수 있었다. 현재까지 Agrobacterium을 이용한 T. harzianum 형질전환은 보고된 바 없다.
Abstract
Effective Agrobacterium-mediated transformation of Trichoderma harzianum could be achieved using the Al_2O_3 particles-abraded mycelia pellets. Transformation efficiency, as percents for the number of hygromcin-resistant mycelia pellets out of total pellets tested, was about 20 in average for Al2O3 experiment. No transformed mycelium was obtained from the intact mycelia pellets. After second round of antibiotics selection, DNA integration of hygromycin resistant gene and the expression of target gene could be confirmed by PCR and RT PCR, respectively. This is the first report of Agrobacterium-mediated T. harzianum transformation
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