Two-step method for constructing Arabidopsis artificial microRNA vectors

被引：0
|
作者：
Xuming Wang
Yong Yang
Jie Zhou
Chulang Yu
Ye Cheng
Chengqi Yan
Jianping Chen
机构：
[1] Zhejiang Academy of Agricultural Sciences,State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, MOA Key Laboratory for Plant Protection and Biotechnology, Zhejiang Provincial Key Laboratory of Plant Virology
来源：
Biotechnology Letters | 2012年 / 34卷
关键词：
artificial miRNA; Gene silencing; Vector construction;
D O I：
暂无
中图分类号：
学科分类号：
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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页码：1343 / 1349
页数：6
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