文献类型: 外文期刊
作者: Wang, Xuming 1 ; Yang, Yong 1 ; Yu, Chulang 1 ; Zhou, Jie 1 ; Cheng, Ye 1 ; Yan, Chengqi 1 ; Chen, Jianping 1 ;
作者机构: 1.Zhejiang Acad Agr Sci, Inst Virol & Biotechnol, MOA, Hangzhou 310021, Zhejiang, Peoples R China; Zhejiang Acad Agr Sci, Zhejiang Prov Key Lab Plant Virol, Hangzhou 310021, Zhejiang, Peoples R China
关键词: Artificial miRNA;Gene silencing;Oryza sativa;Vector construction
期刊名称:MOLECULAR BIOTECHNOLOGY ( 影响因子:2.695; 五年影响因子:2.303 )
ISSN:
年卷期:
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收录情况: SCI
摘要: Artificial microRNA (amiRNA) has become a powerful tool for gene silencing in plants. A new method for easy and rapid construction of rice artificial miRNA vector is described. The procedure involved modification of the pCAMBIA1300-UR vector by insertion of a 'vector modification fragment'. This was prepared from the precursor of Os-amiR528 by eliminating the central miRNA-containing region while simultaneously creating an AfeI restriction site. The fragment was then introduced to the destination vector to produce a multipurpose 'Highly Efficient gene Silencing Compatible vector' (HESC vector). AfeI was used to produce linearized HESC vectors, and a blunt end PCR product that included amiRNA sequence was cloned into this site by a single ligation reaction to create the completed amiRNA vector. Tests showed that the method was highly efficient, and greatly reduced the time needed for vector construction and resulted in a DNA sequence identical to that of the current method, making it particularly suitable for use in a systems biology approach to functional genomic research.
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