Brutnell Thomas P, Conrad Liza J
Boyce Thompson Institute, Cornell University, Ithaca, NY, USA.
Methods Mol Biol. 2003;236:157-76. doi: 10.1385/1-59259-413-1:157.
The transposable element Activator (Ac) has been used in several plant species as a tool for gene isolation and characterization. However, it has not been widely utilized in its native host maize, in part, because of a relatively low germinal transposition rate. The propensity of Ac to move to linked sites provides an opportunity to overcome this limitation when Ac elements are distributed at regular intervals throughout the genome. This chapter details the use of such a system in maize through simple genetic manipulations. A detailed protocol is also provided to clone DNA flanking Ac insertions.
转座元件激活子(Ac)已在多种植物物种中用作基因分离和表征的工具。然而,它尚未在其原生宿主玉米中得到广泛应用,部分原因是其生殖转座率相对较低。当Ac元件在整个基因组中以规则间隔分布时,Ac移动到连锁位点的倾向提供了克服这一限制的机会。本章详细介绍了通过简单的遗传操作在玉米中使用这样一个系统的方法。还提供了一个详细的方案来克隆Ac插入侧翼的DNA。