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。
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.
出处
《植物生理学通讯》
CSCD
北大核心
2010年第7期693-700,共8页
Plant Physiology Communications
