Rigó Gábor, Papdi Csaba, Szabados László
Institute of Plant Biology, Biological Research Center, Szeged, Hungary.
Methods Mol Biol. 2012;913:277-90. doi: 10.1007/978-1-61779-986-0_19.
The controlled cDNA overexpression system (COS) was developed to identify novel regulatory genes in model plants as well as in other species that might have a particular valuable trait. The COS system (Papdi et al. Plant Physiol 147:528-542, 2008) is composed of a random cDNA library prepared in a T-DNA plant expression vector, under the control of the estradiol-inducible XVE promoter. Large-scale genetic transformation of Arabidopsis thaliana generates a transgenic plant population with randomly inserted cDNA clones. Overexpression of the inserted cDNA can create selectable phenotypes, allowing the facile identification and cloning of the responsible genes. Here we describe protocols to create and use the COS system for diverse purposes in plant biology.
可控cDNA过表达系统(COS)的开发是为了在模式植物以及可能具有特定宝贵性状的其他物种中鉴定新的调控基因。COS系统(Papdi等人,《植物生理学》147:528 - 542,2008年)由在T-DNA植物表达载体中制备的随机cDNA文库组成,该文库受雌二醇诱导型XVE启动子的控制。拟南芥的大规模遗传转化产生了一个具有随机插入cDNA克隆的转基因植物群体。插入的cDNA的过表达可以产生可选择的表型,从而便于鉴定和克隆相关基因。在这里,我们描述了在植物生物学中为不同目的创建和使用COS系统的方案。