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Two-step method for constructing Arabidopsis artificial microRNA vectors

文献类型: 外文期刊

作者: Wang, Xuming 1 ; Yang, Yong 1 ; Zhou, Jie 1 ; Yu, Chulang 1 ; Cheng, Ye 1 ; Yan, Chengqi 2 ; Chen, Jianping;

作者机构: 1.Zhejiang Acad Agr Sci, State Key Lab Breeding Base Zhejiang Sustainable, MOA Key Lab Plant Protect & Biotechnol, Zhejiang Prov Key Lab Plant Virol, Hangzhou 310021, Zhejiang, Peoples R China

2.Zhejiang Acad Agr Sci, State Key Lab Breeding Base Zhejiang Sust

期刊名称:BIOTECHNOLOGY LETTERS ( 影响因子:2.461; 五年影响因子:2.457 )

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收录情况: SCI

摘要: Artificial microRNA (amiRNA) technology is used for gene silencing in Arabidopsis. We describe a method for constructing amiRNA vectors that requires only one PCR and one ligation reaction. Vectors produced by this method are the same as those from the method of Schwab et al. (Plant Cell 2006, 18:1121-1133). Transgenic plants created by this method can therefore be tested in the same way or compared with existing transgenic material without the risk of alteration to the amiRNA skeleton. With optimized parameters, 36-42 % colonies had the insertion in the expected orientation and 85-95 % of these had the correct sequence. Using this method, a transient gene knock-down analysis in Arabidopsis could be completed in 4-5 days.

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