Li Tong Xiang, Wang JinKe, Bai YunFei, Sun XiangDong, Lu ZuHong
Chien-Shiung Wu Laboratory, Southeast University, Nanjing 210096, People's Republic of China.
Nucleic Acids Res. 2004 Mar 1;32(4):e45. doi: 10.1093/nar/gnh041.
We report a method called SSH array which combines the suppression subtraction hybridization (SSH) and DNA array techniques to find species-specific DNA probes from genomic DNA (gDNA) for species identification. The method first obtains the differential gDNA fragments between two species by SSH and then hybridizes the differential gDNA fragments with arrays made of multiple whole genomes from several species to screen the unique gDNA fragments for one species. The screened unique gDNA fragments can be used as species-specific probes to differentiate the species they represent from all other species. We used five species of the genus Dendrobrium, D.aurantiacum Kerr, D.officinale Kimura et Migo, D.nobile Lindl., D.chrysotoxum Lindl. and D.fimbriatum Hook., as experimental materials to study the feasibility of the method. The results showed that the method could efficiently obtain different species-specific probes for each of the five species.
我们报道了一种名为SSH阵列的方法,该方法结合了抑制消减杂交(SSH)和DNA阵列技术,从基因组DNA(gDNA)中寻找物种特异性DNA探针用于物种鉴定。该方法首先通过SSH获得两个物种之间的差异gDNA片段,然后将差异gDNA片段与由几个物种的多个全基因组制成的阵列进行杂交,以筛选出一个物种的独特gDNA片段。筛选出的独特gDNA片段可作为物种特异性探针,将它们所代表的物种与所有其他物种区分开来。我们使用了石斛属的五个物种,即金钗石斛、铁皮石斛、鼓槌石斛、密花石斛和流苏石斛作为实验材料来研究该方法的可行性。结果表明,该方法可以有效地为这五个物种中的每一个获得不同的物种特异性探针。