蓖麻毒蛋白A链基因RNAi转化研究  

Construction of RNAi Binary Vector of Ricin A Chain and Its Transformation

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] 

机构地区：[1]内蒙古民族大学,内蒙古通辽028042 [2]内蒙古高校蓖麻产业工程技术研究中心,内蒙古通辽028042

出　　处：《西北植物学报》2013年第1期39-42,共4页Acta Botanica Boreali-Occidentalia Sinica

基　　金：教育部新世纪优秀人才支持计划(NCET-08-0870);内蒙古自然科学基金(2009BS0304)

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762bp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762bp,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。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.

关 键 词：蓖麻毒蛋白 RNAI RNAi表达载体 遗传转化 

分 类 号：Q785[生物学—分子生物学] Q789