摘要
燕窝为宜药宜膳的珍贵药材,其质量与产地和出产环境密切相关。现有的燕窝鉴别方法可在一定程度上鉴别燕窝与其伪品,但无法确定燕窝的基原和产地等质量问题。DNA分子标记技术可以有效地解决这个难题,即从燕窝中得到高质量的DNA以用于后序的PCR和测序。由于燕窝中存在较多内源及外源性污染,其DNA提取技术的难点主要在于摆脱燕窝中的唾液酸糖蛋白干扰和外源性DNA的污染。本文针对燕窝DNA提取方法的优化进行了研究,采用高盐的SDS裂解燕窝样品,氯仿和CTAB去蛋白,低温异丙醇沉淀DNA技术。成功建立了一种简便、高效、稳定的燕窝DNA提取方法。采用本法得到的燕窝DNA可以成功用于燕窝遗传树的建立,而相关燕窝的基原也从而得到了确定。
Edible Bird＇s Nest （EBN） is a kind of precious herbal drug suitable not only for treatment as a drug but also for cooking as a delicious food. The quality of EBN is closely related to its production area and environment, which can not be identified by the current available authentication techniques of EBN that although are capable of differentiating EBN from its counterfeits. However, this problem could be solved by employing the state-of-art DNA molecular marker technique, in which the extraction of high quality DNA from EBN suitable for subsequent PCR and sequencing analyses is the key. As there are a lot of internal and external contaminations in EBN, the key to obtain high quality DNA is therefore to eliminate the interferences from sialoglycoprotein and external DNA contained in EBN. In this work, the extraction method of DNA from EBN was optimized, focusing on eliminating these interferences. Based on this research, a simple, efficient, and stable method for the extraction of DNA from EBN was established using SDS with high concentration of NaCI to lyse the EBN sample, then using chloroform and CTAB to eliminate proteins, and finally using cold isopropyl alcohol to precipitate DNA. Thus obtained DNA has been applied to establish the phylogenic tree of EBN samples, which clearly demonstrates the genetic origin of each EBN sample.
出处
《世界科学技术-中医药现代化》
2010年第2期202-210,共9页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家自然科学基金资助面上项目(K1060015):电泳和mtDNA技术在燕窝鉴别中的应用
负责人:赖小平
关键词
燕窝
DNA
提取方法
Edible Bird＇s Nest, DNA, Extraction method
