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目的 :建立中药红曲基原菌的随机扩增多态性 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 ,PHYL IP 3.5 c进行聚类分析。 结果 :从 6 0个随机引物中筛选出 S2 82 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。
Objective : To establish a random amplified polymorphic DNA (RAPD) method for the analysis of the original genus Monascus. METHODS: Genomic DNA of 7 Monascus species such as Monascus purpureus and 1 Aspergillus terreus were extracted with CTAB method; ethidium bromide fluorescence intensity and spectrophotometry were used to determine DNA concentration; agarose gel electrophoresis was used to detect PCR amplification products. , PHYL IP 3.5 c for cluster analysis. RESULTS: More than 10 primers including S2 82 were screened out from 60 random primers with good characteristics. The inter-species fingerprints of Monascus spp. showed large differences and showed obvious DNA amplification product polymorphisms. . Conclusion : RAPD analysis technology can be used as one of the objective and effective methods of fungal system classification.