| 蓖麻毒蛋白A链基因RNAi转化研究 |
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| 引用本文： | 陈永胜,王永佳,王 莹,黄凤兰,李国瑞,王文跃.蓖麻毒蛋白A链基因RNAi转化研究[J].西北植物学报,2013,33(1):39-42. |
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| 作者姓名： | 陈永胜 王永佳 王 莹 黄凤兰 李国瑞 王文跃 |
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| 作者单位： | 1. 内蒙古民族大学,内蒙古通辽028042;内蒙古高校蓖麻产业工程技术研究中心,内蒙古通辽028042
2. 内蒙古民族大学,内蒙古通辽,028042 |
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| 基金项目： | 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） |
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| 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.
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| 关 键 词： | 蓖麻毒蛋白 RNAi RNAi表达载体 遗传转化 |
Construction of RNAi Binary Vector of Ricin A Chain and Its Transformation |
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| CHEN Yongsheng,WANG Yongji,WANG Ying,HUANG Fenglan,LI Guorui,WANG Wenyue.Construction of RNAi Binary Vector of Ricin A Chain and Its Transformation[J].Acta Botanica Boreali-Occidentalia Sinica,2013,33(1):39-42. |
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| Authors: | CHEN Yongsheng WANG Yongji WANG Ying HUANG Fenglan LI Guorui WANG Wenyue |
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| Institution: | 1(1 Inner Mongolia University for Nationalities,Tongliao,lnner Mongolia 028042,China;2 Engineering Research Center of Castor at Universities of lnner Mongolia Autonomous,Tongliao,lnner Mongolia 028042,China) |
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| 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. |
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| Keywords: | Ricin RNA interference binary expression vector genetic transformation |
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