| 人工microRNA干扰拟南芥AtCDKC;1和AtCDKC;2基因表达的初步研究 |
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| 引用本文： | 赵臻,邱凯,蒯本科.人工microRNA干扰拟南芥AtCDKC;1和AtCDKC;2基因表达的初步研究[J].植物生理学通讯,2010(7):693-700. |
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| 作者姓名： | 赵臻 邱凯 蒯本科 |
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| 作者单位： | 复旦大学生命科学学院 遗传工程国家重点实验室和植物科学研究所,上海200433 |
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| 摘 要： | 以来源于拟南芥的microRNA序列为骨架,构建抑制AtCDKC;1和AtCDKC;2基因的人工microRNAs载体,研究其对目的基因表达的抑制效果。选择AtCDKCs基因的特异性序列,通过重叠PCR的方法改造拟南芥microRNA164a骨架序列,连接到双元载体pPZPY122,在农杆菌介导下转化拟南芥。RT-PCR分析表明,人工microRNA能够显著抑制目的基因的表达,获得了抑制效果明显的转基因植株,并且对AtCDKCs在拟南芥生长发育中的作用进行了初步的研究。
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| 关 键 词： | 周期蛋白依赖性激酶 人工microRNA RNA干扰 基因表达 拟南芥发育 |
A Preliminary Analysis of Artificial MicroRNAs-Mediated Interference of AtCDKC;1 and AtCDKC;2 Expression in Arabidopsis |
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| ZHAO Zhen,QIU Kai,KUAI Ben-Ke.A Preliminary Analysis of Artificial MicroRNAs-Mediated Interference of AtCDKC;1 and AtCDKC;2 Expression in Arabidopsis[J].Plant Physiology Communications,2010(7):693-700. |
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| Authors: | ZHAO Zhen QIU Kai KUAI Ben-Ke |
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| Institution: | State Key Laboratory of Genetic Engineering and Institute of Plant Biology,School of Life Sciences,Fudan University,Shanghai 200433,China |
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| Abstract: | By using Arabidopsis endogenous microRNA sequences,artificial microRNAs （amiRNAs） were generated to knock down the expressions of AtCDKCs.To construct plasmids,oligonucleotide sequences targeting AtCDKCs-specific locus were used to engineer microRNA164a by overlapping PCR and the resultant sequence was inserted into the binary vector pPZPY122.Arabidopsis of wild type （Col-0） was transformed by the Agrobacterium-mediated approach.RT-PCR analysis showed that the target genes were both specifically and efficiently silenced in transgenic lines.The roles of AtCDKCs in the growth and development of Arabidopsis was analyzed based on the transgenic plants. |
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| Keywords: | AtCDKC artificial microRNA RNA interference gene expression Arabidopsis development |
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