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蔊菜病毒诱导基因沉默体系构建

DOI: 10.7668/hbnxb.2013.06.011, PP. 65-70

Keywords: 病毒诱导的基因沉默,蔊菜,CLA基因,烟草脆裂病毒

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Abstract:

构建蔊菜的病毒诱导基因沉默(Virusinducedgenesilencing,VIGS)体系,旨在建立蔊菜的快速基因功能验证的方法。通过RT-PCR扩增出棉花、大豆、拟南芥的1-脱氧木酮糖-5-磷酸合成酶(Cloroplastoalterados,CLA)基因片段,并将其分别构建到烟草脆裂病毒(Tobaccorattlevirus,TRV)介导的VIGS载体,通过农杆菌GV3101侵染蔊菜并观察蔊菜的表型变化。结果显示,3种不同的CLA基因所侵染的蔊菜均出现不同程度的白化现象,半定量RT-PCR检测显示CLA的mRNA被显著降解,表明成功构建了蔊菜的VIGS体系。

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