| 甘蓝型油菜异三聚体G蛋白原生质体瞬时表达体系的建立 |
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| 引用本文： | 陈云,于一帆,潘淑芬,周研,葛会敏,刘立军.甘蓝型油菜异三聚体G蛋白原生质体瞬时表达体系的建立[J].华北农学报,2016(6):21-25. |
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| 作者姓名： | 陈云 于一帆 潘淑芬 周研 葛会敏 刘立军 |
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| 作者单位： | 扬州大学江苏省作物遗传生理重点实验室,江苏扬州,225009 |
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| 基金项目： | 国家自然科学基金项目(31371562 |
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| 摘 要： | 为建立甘蓝型油菜异三聚体G蛋白各组分原生质体瞬时表达体系,以油菜叶片为试材,从试材选取、平摇时间和聚乙二醇浓度方面优化了油菜原生质体制备条件和转化条件。结果表明,选取30 d苗龄叶片、平摇10 min时原生质体游离状态最佳,产量为10.73×10~6/m L,活力可达96.4%。采用30%PEG转化原生质体时,目的蛋白表达量最高。Western Bolt的检测分析表明,异三聚体G蛋白各组分均参与了甘蓝型油菜对ABA的应答反应,当ABA浓度分别为100,150,100,50 mmol/L时,Bn GA1、Bn GB1、Bn GG2和BnRGS1蛋白的表达量最高。
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| 关 键 词： | 油菜 原生质体瞬时表达 异三聚体G蛋白 |
Establishment of Heterotrimeric G Protein Gene Protoplast Transient Expression System in Brassica napus |
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| Abstract: | To establish the protoplast transient expression system of the components of heterotrimeric G protein in Brassica napus,the key factors in protoplast preparation and transformation including explant,flat rolling time and polyethylene glycol(PEG) concentration were optimized.The results showed that the best yield and viability of protoplasts were 10.73 × 106/mL and 96.4％ respectively,with the optimum conditions as 30 days seedling age and 10 min rolling time.The expression of target proteins reached the maximum with 30％ PEG.The responses of G protein to ABA were investigated using the protoplast transient system by Western Blot detection.The highest expression levels of BnGA1,BnGB1,BnGG2 and BnRGS1 were observed when the ABA concentration was 100,150,100,50mmol/L,respectively. |
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| Keywords: | Brassica napus Protoplast transient expression Heterotrimeric G protein |
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