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中间锦鸡儿fad2基因片段克隆、反义表达载体构建及遗传转化初步研究

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Keywords: 中间锦鸡儿,生物柴油,fad2基因,反义表达载体,遗传转化

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Abstract:

以我国重要的生物能源灌木―――中间锦鸡儿枝叶和种子为材料,根据GenBank中已经发表fad2基因的同源序列,利用PCR技术克隆得到基因片段。在GenBank中Blast (GenBank登录号AY957393)所得基因片段和同属豆科的Glycine max Gmfad2-2a同源性高达88% ,位于fad2基因编码区中部。将所得片段经BamH I和Sac I酶切后插入表达载体质粒pB I121,构建了反义表达载体pB I121fad2,并利用农杆菌介导法转入烟草叶片,获得了抗卡那霉素和氨苄青霉素的再生烟草植株。初步分析结果表明:与对照烟草相比,转基因烟草种子脂肪酸含量没有明显差异,而亚油酸则减少10. 3%。

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