植物基因启动子的克隆方法及其应用

植物基因的表达调控已成为分子生物学研究热点,启动子是基因表达调控的重要顺式元件,启动子的克隆对于研究基因表达调控、构建基因工程载体、表达目的蛋白有着重要的意义。启动子克隆的方法很多,从常用的启动子陷阱技术筛选启动子到PCR方法的应用,此后相继问世的一些基于PCR的克隆启动子技术,如载体锚定PCR、反向PCR、接头PCR、交错热不对称PCR等,为克隆启动子提供了更可靠,更合理的方法。本文着重介绍了几种植物基因启动子的克隆方法,分析了它们的优缺点,并展望了今后的研究前景。

Cloning Methods of Plant Gene Promoters and Their Applications

The expression and regulation of plant gene has been the hotspot study in molecular biology. Promoter is an important cis-acting element. Cloning promoter is important to study gene regulation, vectors construction and target proteins expression. From the frequently used way that applying promoter trapping for choosing promoter to the using of PCR, there were lots of ways for promoter cloning. Afterwards, a series of techniques based on PCR for promoter cloning such as A-PCR, I-PCR, LA-PCR, TAIL-PCR were developed in succession. They offered more reliable and reasonable ways of cloning the promoter. Some ways of cloning plant gene promoters were introduced, meanwhile the limitation of these ways was introduced and their developing prospect was also discussed in this paper.