高番茄红素加工番茄品系的RAPD引物筛选及特异片段的克隆
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甘肃省农业生物技术研究与应用项目(GNSW-2006-02)


Selection of RAPD Marker for High Lycoppene in Tomato and Its Specific Fragment Cloning
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    摘要:

    以加工番茄高茄红素近等基因系为材料,分别从高茄红素含量及低茄红素含量植株中提取DNA构建DNA池,采用RAPD技术,从260个随机引物中筛选出1个在2基因池间具有特异性的引物S636,在轮回亲本及回交后代单株的DNA中进行验证,证明了该引物扩增出的特异性片段是1个与番茄耐高茄红素含量连锁的RAPD标记.经琼脂糖凝胶电泳回收扩增出的特异条带与载体pMD-18T连接,并转入大肠埃希菌DH-5α,对克隆片段测序结果表明其实际大小为827 bp,这为转化成稳定的SCAR标记奠定了基础.

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    The near isogenic lines of high and low lycoppene processing tomato were used as experiment materials; DNA was extracted from these plants to develop high and low lycoppene DNA gene pools, respectively. RAPD techniques were adopted to screen molecular marker linked to high lycoppene, and only primer S636 was found to be polymorphic between near isogenic lines of high and low lycoppene processing tomato from 260 RAPD random primers. It had been proved that a specific fragment of RAPD marker was linked to high lycoppene in the back cross parent and the single plants in the backcross population. The specific fragment was extracted from the agarose and ligated with pMD-18T vector, then transformed into DH-5α. The positive clone was sequenced to find a 827 bp fragment. This fragment could be used to develop SCAR marker for steady lycoppene.

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谢 放,张 楠.高番茄红素加工番茄品系的RAPD引物筛选及特异片段的克隆[J].华南农业大学学报,2012,33(4):503-506

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  • 收稿日期:2012-02-16
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  • 在线发布日期: 2012-11-16