Optimization of Particle Bombardment Conditions by Monitoring of Transient sGFP(S65T) Expression in Transformed Soybean
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概要
- 論文の詳細を見る
In an attempt to increase the efficiency of soybean transformation by particle bombardment, we examined the effects of bombardment parameters such as gold particle size, target distance, acceleration pressure, amount of DNA per bombardment, and number of bombardments. Transgene delivery to embryogenic tissue grown in suspension culture was evaluated by monitoring the transient expression of a gene for a modified form of jellyfish green fluorescent protein [sGFP(S65T)]. Optimal transient expression of sGFP(S65T) was obtained when the tissue was bombarded twice at an acceleration pressure of 7.6MPa (1,100psi) and a distance of 6cm with gold particles that were 0.6μm in diameter and coated with 0.8μg of DNA. Application of these optimized conditions proved effective for the generation of stable transgenic soybean plants. Stable transgene integration in the transformants was confirmed by Southern blot analysis. The average transformation efficiency achieved with the optimized protocol was siginificantly higher than that with the conventional protocol.
- 日本育種学会の論文
著者
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Wakasa Kyo
Core Research Of Science And Technology(crest):national Institute Of Crop Science:tokyo University O
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ISHIMOTO Masao
National Agriculture Research Center
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Khalafalla Mutasim
National Agricultural Research Center For Western Region:core Research Of Science And Technology(cre
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El Shemy
National Agricultural Research Center For Western Region:core Research Of Science And Technology(cre
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Nakamoto Yumi
National Agricultural Res. Center For Hokkaido Region
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Ishimoto Masao
National Agricultural Res. Center For Hokkaido Region
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Rahman Shaikh
Present Address: National Agricultural Res. Center For Western Region
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Rahman Shaikh
National Agricultural Research Center for Western Region
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Nakamoto Yumi
Core Research of Science and Technology(CREST)
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Ishimoto Masao
National Institute Of Agrobiological Sciences
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Ishimoto Masao
National Inst. Of Agrobiological Sciences
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El-Shemy Hany
National Agricultural Research Center for Western Region
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