Martienssen R A
Cold Spring Harbor Laboratory, Box 100, Cold Spring Harbor, NY 11724, USA.
Proc Natl Acad Sci U S A. 1998 Mar 3;95(5):2021-6. doi: 10.1073/pnas.95.5.2021.
Transposable elements provide a convenient and flexible means to disrupt plant genes, so allowing their function to be assessed. By engineering transposons to carry reporter genes and regulatory signals, the expression of target genes can be monitored and to some extent manipulated. Two strategies for using transposons to assess gene function are outlined here: First, the PCR can be used to identify plants that carry insertions into specific genes from among pools of heavily mutagenized individuals (site-selected transposon mutagenesis). This method requires that high copy transposons be used and that a relatively large number of reactions be performed to identify insertions into genes of interest. Second, a large library of plants, each carrying a unique insertion, can be generated. Each insertion site then can be amplified and sequenced systematically. These two methods have been demonstrated in maize, Arabidopsis, and other plant species, and the relative merits of each are discussed in the context of plant genome research.
转座元件为破坏植物基因提供了一种便捷灵活的手段，从而能够对其功能进行评估。通过对转座子进行工程改造，使其携带报告基因和调控信号，可以监测并在一定程度上操纵靶基因的表达。本文概述了利用转座子评估基因功能的两种策略：第一，聚合酶链式反应（PCR）可用于从大量诱变个体群体中鉴定出携带特定基因插入的植物（位点选择转座子诱变）。该方法要求使用高拷贝转座子，并且需要进行相对大量的反应以鉴定感兴趣基因中的插入情况。第二，可以构建一个大型植物文库，每个植物携带一个独特的插入片段。然后可以系统地扩增并测序每个插入位点。这两种方法已在玉米、拟南芥和其他植物物种中得到验证，并且在植物基因组研究的背景下讨论了每种方法的相对优点。
