A Golden Gate and Gateway double-compatible vector system for high throughput functional analysis of genes
文献类型： 外文期刊
作者：
Luo, Yanjie
1
;
Qiu, Yang
1
;
Na, Ren
3
;
Meerja, Farida
1
;
Lu, Qing Shi
1
;
Yang, Chunyan
3
;
Tian, Lining
1
;
作者机构： 1.Agr & Agri Food Canada, London Res & Dev Ctr, London, ON N5V 4T3, Canada
2.Chinese Acad Agr Sci, Inst Vegetables & Flowers, Beijing 100081, Peoples R China
3.Hebei Acad Agr & Forestry Sci, Inst Cereal & Oil Crops, Shijiazhuang 050035, Hebei, Peoples R China
4.Western Univ, Dept Biol, London, ON N6A 5B7, Canada
关键词： Golden Gate assembly; Gateway cloning; BiFC; Artificial microRNA; CRISPR/Cas9
期刊名称：PLANT SCIENCE （ 影响因子：4.729； 五年影响因子：5.132 ）
ISSN： 0168-9452
年卷期： 2018 年 271 卷
页码：
收录情况： SCI
摘要： A major research topic nowadays is to study and understand the functions of the increasing number of predicted genes that have been discovered through the complete genome sequencing of many plant species. With the aim of developing tools for rapid and convenient gene function analysis, we have developed a set of "pGate" vectors based on the principle of Golden gate and Gateway cloning approaches. These vectors combine the positive aspects of both Golden gate and Gateway cloning strategies. pGate vectors can not only be used as Golden gate recipient vectors to assemble multiple DNA fragments in a pre-defined order, but they can also work as an entry vector to transfer the assembled DNA fragment(s) to a large number of already-existing, functionally diverse, Gateway compatible destination vectors without adding additional nucleotides during cloning. We show the pGate vectors are effective and convenient in several major aspects of gene function analyses, including BiFC (Bimolecular fluorescence complementation) to analyze protein-protein interaction, amiRNA (artificial microRNA) candidate screening and as assembly of CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeats, CRISPR-associated protein-9 nuclease) system elements together for genome editing. The pGate system is a practical and flexible tool which can facilitate plant gene function research.
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