GUS 유전자를 이용한 Agrobacterium 매개 Siberian Wildrye Grass의 형질전환 효율에 미치는 영향
Optimization of Agrobacterium-Mediated Transformation Parameters for Siberian Wildrye Grass Callus using GUS Gene as a Marker
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참고문헌
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초록
분자육종기술을 이용하여 불량 환경조건에서도 강한 형질전환 목초를 개발하기 위한 기초실험으로 몽골지역에 자생하는 Siberian wildrye grass를 이용하여 Agrobacterium을이용한 형질전환에 미치는 몇 가지 요인들을 조사하였다. Agrobacterium 감염 시에 Agrobacterium strain EHA105을 이용하여 100 μM의 acetosyringone(AS)을 첨가해 주었을 때형질전환 효율이 증가되었으며, 공동배양기간을 5일까지 증가시켰을 때 형질전환효율이 가장 높게 나타났다. 본 연구에서 GUS 유전자를 이용한 형질전환의 최적 조건은 건조와염해에 강한 내성을 부여하는 유전자(DREB) 도입에 유용하게 이용될 것이다.
Abstract
Gene manipulation of Siberian wildrye grass through transgenic technology can modify grass feature for further improvement in the forage production. In order to optimize transformation conditions of Siberian wildrye grass, the effects of transformation efficiency was investigated with GUS gene. Seed-derived calli were infected and co-cultured with Agrobacterium tumefaciens carrying the binary vector pCAMBIA3301 encoding the GUS gene in the T-DNA region. The effects of several factors on transformation and the expression of the GUS gene were investigated. The highest transformation efficiency was obtained when embryogenic calli were inoculated with Agrobacterium strain EHA101 in the presence of 200 μM acetosyringone and coculture for 5days. The present study was framed and materialized to standardize an efficient, high frequency and reproducible genetic transformation protocol for aromatic Siberian wildrye grass using Agrobacterium tumefaciens.
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