微型反向重复转座元件(MITE)靶区域扩增多态性:一种基于MITE的分子标记方法在水稻及其他植物上的应用

MITE-TRAP:A Marker Technique Based on Miniature Inverted Repeat Transposable Element and Target Region Ampilifcation Polymorphism for Rice and Other Plants

利用根据水稻微型反向重复转座元件（miniature inverted repeat transposable element，MITE）序列设计的特异引物，以及靶区域扩增多态性（target region amplification polymorphism，TRAP）方法中的随机引物及扩增程序对不同的水稻材料进行PCR扩增来检测MITE侧翼为基因区域的多态性，称之为微型反向重复转座元件靶区域扩增多态性（MITE-TRAP）。每次扩增能产生1条或多条清晰条带，条带的大小为100～1500bp，可在1．5％的琼脂糖凝胶上分离，有较好的可重复性，并发现了不同水稻品种间的多态性条带。进一步用基于水稻预测的MITE序列设计的特异引物对来自棉花、番茄及拟南芥的DNA样品进行MITE-TRAP扩增，均能成功扩增出条带，显示这种方法可以直接在其他植物中应用。还进一步讨论了该方法的优点及其可能的应用。

A MITE-TRAP marker method was developed, in which the fixed primer, designed from the miniature inverted repeat transposable element（MITE） sequence, was instead of EST or gene sequence in target region amplification polymorphism （TRAP） technique, and the arbitrary primer was the same as that in TRAP. As different rice materials were analyzed by using the fixed primer designed from the MITE sequence in rice, each PCR reaction could amplify one or several clear bands with sizes ranging from 100-1500 bp on a 1.5 %agarose gel, which possessed better reproducibility. Some polymorphic bands in these rice materials were also obtained. Moreover, this method could be applied in other plant species directly, such as cotton,tomato and arabidopsis. Some advantages of the method and its application were discussed.