一种中药鉴定gDNA芯片的制备与展望

中药鉴定是中药质量控制系统的首要环节,中药鉴定技术的不断发展提高了药材品质评价水平。至今,中药鉴定技术经历着从传统生药鉴定学的细胞和亚细胞水平向遗传物质DNA分子水平发展的历程,运用DNA芯片技术进行中药鉴定成为中药鉴定发展的方向之一。介绍了筛选gDNA种特异性探针的抑制性差减微阵列“SSH-ar-ray”新技术,该技术是将抑制性差减杂交与基于尼龙膜微阵列杂交相结合。以名贵中药材石斛5个品种作为实验材料,首先是用抑制性差减杂交获得2个品种之间的gDNA差异片段,然后用这些gDNA差异片段与由所有研究品种的gDNA制备成的微阵列杂交筛选到某一个品种的种特异性gDNA片段。这些种特异性gDNA探针作为种特异性探针进行物种鉴定。在此基础上建立中药鉴定gDNA芯片的方法并对其应用进行了展望探讨。

Preparation and prospect of gDNA microarray for species identification of Chinese materia medica

The identification of traditional Chinese herbs(TCH) is very important in the quality control system of TCH.The development of the TCH identification techniques has improved the quality evaluation level of TCH.By so far,the TCH identification techniques have been developed from traditional pharmacognostic identification methods based on cell and sub-cell to genetic DNA(gDNA) level.It is a tendency to use DNA microarray to identify the species of TCH in recent years.In this review,a new method called suppression subtraction hybridization and DNA array(SSH-array) was introduced,which combined the SSH technique with microarray hybridization based on nylon film.Five plants from Dendrobium Sw.were used as experimental materials.Firstly the differential gDNA fragments between two species by SSH-array were obtained and then the diferential gDNA fragments with the microarrays made up of multiple whole genomes from several species were hybridized to screen the uniqued gDNA fragments for one species.The screened unique gDNA fragment can be used as species-specific probes for identifying the species.Based on the method,a procedure for species identification of TCH was established with gDNA microarray and its applied prospects were discussed.