转化(遗传学)
可选择标记
生物
农杆菌
根癌农杆菌
卡那霉素
清脆的
基因
遗传学
子叶
基因组
Cas9
计算生物学
植物
作者
Joyce Van Eck,Patricia Keen,Michelle Tjahjadi
出处
期刊:Methods in molecular biology
日期:2018-11-11
卷期号:: 225-234
被引量:92
标识
DOI:10.1007/978-1-4939-8778-8_16
摘要
Tomato is both an important food crop and serves as a model plant species that is used for various research investigations including understanding gene function. Transformation is commonly utilized to facilitate these investigations in combination with all the extensive genetic and genomic resources available for tomato. The transformation protocol routinely used in our laboratory has been applied to many different tomato genotypes and relies on Agrobacterium tumefaciens infection of young cotyledon sections. We have used vector systems for overexpression, RNA interference for gene silencing, and CRISPR/Cas9 for genome editing. Vectors used to design gene constructs contained selectable marker genes that conferred resistance to kanamycin, hygromycin, and the herbicide component, bialaphos. The protocol we follow for Agrobacterium-mediated transformation of both cultivated and wild species of tomato is detailed in this chapter.
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