随机扩增多态性DNA技术在中药红曲基原菌系统分类中的应用

目的：建立中药红曲基原菌的随机扩增多成性ＤＮＡ（ＲＡＰＤ）分析方法。方法：以ＣＴＡＢ法提取紫色红曲霉等７种红曲霉和１株土曲霉的基因组ＤＮＡ；溴化乙啶荧光强度和分光光度双重测定法确定ＤＮＡ浓度；琼脂糖凝胶电泳法检测ＰＣＲ扩增产物，ＰＨＹＬＩＰ３．５ｃ进行聚类分析。结果：从６０个随机引物中筛选出Ｓ２８２等１０个引物的扩增产物谱带多，特征好；红曲霉属种间指纹图谱差异较大，呈现出明显的ＤＮＡ扩增产物多态性。结论：ＲＡＰＤ分析技术可作为真菌系统分类的客观而有效的手段之一。

Application of random amplified polymorphism DNA technique in systematic classif ication of origin fungi of Chinese traditional medicine Red Kojic

Objective:To establish a random amplified polymorphism DNA(RAPD) method for classification of origin fungi of the Chinese traditional medicine Red Kojic. Methods:The genome DNA of 7 ordinary species Monascus including M. purpureus and a strain of Aspergillus terreus were extracted by CTAB. The content of DNA was assayed doubly by fluorescence intensity of ethidium bromide (EB) and spectrometer. Amplification products were detected by agarose gel electrophoresis and clustering analysis by PHYLIP 3.5c. Results:A characteristic pattern was produced depend on one of the 10 primers screened from 60 random primers, and the genome was amplified. The diversities of the fingerprint patterns of Monascus was obvious, by which Monascus can be distinguished easily. The results were consistent with that of morphologic study. Conclusion:RAPD is a new assay technique. It can be used to classify and identify of Monascus fungi.