东方山羊豆GoMIPS双元表达载体的构建及鉴定
Acta Ecologiae Animalis Domastici（2013）
Abstract
从东方山羊豆(Galega.orientalis L.)中克隆出的GoMIPS(肌醇-1-磷酸合成酶)基因序列设计合成1对带有NcoI和SpeI酶切位点的引物,用RT-PCR方法从东方山羊豆幼嫩叶片总RNA中扩增出GoMIPS基因的cDNA序列,经纯化后,克隆至pMD19-T载体上,构建pMD-MIPS的中间载体,测序鉴定。然后用NcoI和SpeI限制性酶对含有目的基因的pMD-MIPS和pCAMBIA1302空载体进行双酶切,通过酶切鉴定和测序分析表明,已成功构建了CaMV35S启动子驱动GFP报告基因的双元植物表达载体pCAMBIA1302-MIPS。
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Key words
pCAMBIA1302 vector,GoMIPS(inositol-1-phosphate synthase),Galega.orientalis L.,pMD19-T vector
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