Ujino T, Kawahara T, Tsumura Y, Nagamitsu T, Yoshimaru H, Ratnam W
Bio-technology Division, Forestry and Forest Products Research Institute, Ibaraki, Japan.
Heredity (Edinb). 1998 Oct;81 ( Pt 4):422-8. doi: 10.1046/j.1365-2540.1998.00423.x.
Nine simple sequence repeat (SSR) markers were developed from Shorea curtisii using two different methods. One SSR locus was isolated by the commonly used method of screening by colony hybridization, and the other eight loci were isolated by a vectorette PCR method. Primer pairs were designed based on the sequences of all these SSR loci. Analysis of 40 individuals of S. curtisii from natural forest in Malaysia revealed that all SSR loci were polymorphic. Four SSR markers, Shc01, Shc04, Shc07 and Shc09, were highly polymorphic. We have also tested the applicability of these SSR printers to other species of Dipterocarpaceae using PCR amplification. Because the flanking region sequences of the S. curtisii SSRs were well conserved within this family, the SSR primers for S. curtisii can be applied to almost all species of Dipterocarpaceae.
利用两种不同方法从柯氏娑罗双(Shorea curtisii)中开发了9个简单序列重复(SSR)标记。一个SSR位点通过常用的菌落杂交筛选方法分离,另外8个位点通过载体PCR方法分离。根据所有这些SSR位点的序列设计了引物对。对来自马来西亚天然林的40个柯氏娑罗双个体进行分析,结果表明所有SSR位点均具有多态性。四个SSR标记,即Shc01、Shc04、Shc07和Shc09,具有高度多态性。我们还通过PCR扩增测试了这些SSR引物对龙脑香科其他物种的适用性。由于柯氏娑罗双SSRs的侧翼区域序列在该科内保守性良好,因此柯氏娑罗双的SSR引物几乎可应用于龙脑香科的所有物种。