---

作 者 ：陈永胜1,2，王永佳1，王 莹1，黄凤兰1,2，李国瑞1,2，王文跃1

期 刊 ：西北植物学报 2013年 1期 页码：39~42

关键词：蓖麻毒蛋白；RNAi；RNAi表达载体；遗传转化；

Keywords:Ricin, RNA interference, binary expression vector, genetic transformation,

摘 要 ：通过基因沉默技术调控蓖麻毒蛋白A链基因的表达，以期获得低毒蓖麻新材料。利用基因克隆技术获得蓖麻毒蛋白A链基因762 bp片段,命名为RTA基因。进一步利用该基因构建了植物RNAi表达载体pBI-RTA-S-AS,通过农杆菌介导法转化蓖麻子叶节,用卡那抗性筛选转化再生植株，PCR进一步鉴定转基因植株。结果表明：克隆得到目的基因长762 bp，与预期结果一致；卡那抗性筛选和PCR鉴定结果显示，获得了3株转基因阳性植株。

Abstract:In order to obtain a new castor with low toxity,RNAi interference technology was used to regulate the expression of the RTA(ricin toxin A-subunit) gene.The RTA gene are amplified by Polymerase Chain Reaction technology,and then RTA transgenic expression vector(pBI-RTA-S-AS) was constracted successfully.Secondly,pBI121-RTA-dsRNA vector was transferred into cotyledonary node by Agrobacterium tumefaciens-mediated transformation.Transformed castor plants were identified by growth on medium containing 25mg/L Kanamycin and PCR.The results showed that 762 bp fragment was obtained.Three transformants were obtained and all of them were further identified as positive transgenitic plants by PCR.

---