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Author:
Date:
2014-12-20
[Abstract] This is a protocol to produce stable silencing efficacy and efficiency for VIGS using CymMV as a silencing vector for floral functional genomics in Phalaenopsis orchids. This protocol is established based on a method created by Lu et al. (2007), and then modified by Hsieh et al. (2013a; 2013b), Lu et al. (2012) successfully engineered a cloning vector (pCymMV-Gateway) in that the target gene fragment is simple to insert and can be manipulated with high efficiency. The silencing vector is inoculated into plants by Agro-inoculation by using Agrobacterium tumefaciens (A. tumefaciens) strain EHA105. Agro-infiltration of leaves for use in VIGS study of orchid flowers is a time saver and produces less damage to flower buds.
[摘要] 这是使用CymMV作为蝴蝶兰花中的花功能基因组的沉默载体产生VIGS的稳定沉默效率和效率的方案。 该协议是基于由Lu等人（2007）创建的方法建立的，然后由Hsieh等人（2013a; 2013b）修改， （2012）成功地设计了克隆载体（pCymMV-Gateway），其中靶基因片段易于插入并且可以高效率操作。 通过使用根癌土壤杆菌（根瘤土壤杆菌）菌株EHA105通过农杆菌接种将沉默载体接种到植物中。 用于VIGS研究兰花花的叶的农杆菌浸润是一种节省时间并且对花芽产生较少的损害。
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