何首乌种特异DNA探针的筛选

Screening of species-specific DNA probes for the identification of Fallopia multiflora

为筛选得到何首乌(Fallopia multiflora)种特异DNA探针,通过何首乌gDNA和其近缘植物毛脉蓼(F.multiflora var.cillinerve)gDNA之间的的抑制消减杂交(Suppression subtraction hybridization,SSH)得到何首乌的差减片段,再将标记的差减片段和多物种gDNA阵列杂交筛选得到了4条何首乌DNA探针,探针通过反向斑点杂交检测,能很好地区分何首乌及其混伪品。获得的探针在中药鉴定基因芯片的制备及何首乌种质和生药鉴定方面具应用前景。

To find specie-specific DNA probes of Fallopia multiflora, the genomic DNA （gDNA） suppression subtraction hybridization （SSH） between Fallopia multiflora and F. multiflora var. ciliinervis was firstly performed. The obtained differential gDNA fragments by SSH were then hybridized with gDNA arrays made of multiple whole genomes of several species （adulterants and/or closely related plants of F. multiflora） and four of the differential fragments were screened unique representing F. multiflora, which can be used as F. multifiora speciss-specific probes. The DNA probes were tested by reserved dot blot hybridization, the results demonstrated the probes could be reliably used to identify F. multiflora. The study of this paper offers the probes representing F. multiflora. The obtained probes are capable to be applied on germplasm or crude drug authentication of F. multiflora and the preparation of Chinese traditional medicines identification gene chip.