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草业学报  2009 

豌豆清蛋白1(PA1)基因的克隆及对苜蓿的转化

, PP. 117-125

Keywords: 豌豆清蛋白(PA1)基因,富硫氨基酸,苜蓿,遗传转化

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Abstract:

本研究用PCR方法从豌豆(食荚大菜豌)克隆出富含硫氨基酸、同时具有抗虫作用的双功能的蛋白质基因-豌豆清蛋白1(PA1)基因,并构建了植物表达载体pCAMBIAl301-PA1。采用农杆菌介导法转化了紫花苜蓿,并对其转化体系进行了优化,得到了多个转基因胚性愈伤组织及其再生植株。PCR和Southern杂交检测表明,PA1基因和潮霉素抗性基因已被整合到了宿主细胞。SDS-PAGE分析表明该基因在再生植株中有一定表达。游离氨基酸分析表明,PA1基因的表达转基因苜蓿中蛋氨酸和半胱氨酸的含量从0.1%提高到0.4%。

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