Lian Chunlan L, Abdul Wadud Md, Geng Qifang, Shimatani Kenichiro, Hogetsu Taizo
Asian Natural Environmental Science Center, The University of Tokyo, 1-1-8 Midori-cho, Nishitokyo, Tokyo, 188-0002, Japan,
J Plant Res. 2006 Jul;119(4):415-7. doi: 10.1007/s10265-006-0274-2. Epub 2006 Apr 25.
An approach for developing codominant polymorphic markers (compound microsatellite (SSR) markers), with substantial time and cost savings, is introduced in this paper. In this technique, fragments flanked by a compound SSR sequence at one end were amplified from the constructed DNA library using compound SSR primer (AC)6(AG)5 or (TC)6(AC)5 and an adaptor primer for the suppression-PCR. A locus-specific primer was designed from the sequence flanking the compound SSR. The primer pairs of the locus-specific and compound SSR primers were used as a compound SSR marker. Because only one locus-specific primer was needed for design of each marker and only a common compound SSR primer was needed as the fluorescence-labeled primer for analyzing all the compound SSR markers, this approach substantially reduced the cost of developing codominant markers and analyzing their polymorphism. We have demonstrated this technique for Dendropanax trifidus and easily developed 11 codominant markers with high polymorphism for D. trifidus. Use of the technique for successful isolation of codominant compound SSR markers for several other plant species is currently in progress.
本文介绍了一种开发共显性多态性标记（复合微卫星（SSR）标记）的方法，该方法可大幅节省时间和成本。在这项技术中，使用复合SSR引物（AC）6（AG）5或（TC）6（AC）5以及用于抑制PCR的接头引物，从构建的DNA文库中扩增一端侧翼为复合SSR序列的片段。根据复合SSR侧翼的序列设计位点特异性引物。位点特异性引物和复合SSR引物的引物对用作复合SSR标记。由于每个标记的设计只需要一个位点特异性引物，并且只需要一个通用的复合SSR引物作为分析所有复合SSR标记的荧光标记引物，因此该方法大大降低了开发共显性标记及其多态性分析的成本。我们已将该技术应用于裂叶山参，并轻松开发出11个具有高多态性的裂叶山参共显性标记。目前正在将该技术用于成功分离其他几种植物物种的共显性复合SSR标记。
