Ding Hong-Mei, Ding Zhi-Shan, Li Hai-Bo, Chen Ni-Pi
Analytical Testing Center, Zhejiang Chinese Medical University, Hangzhou 310053, China.
Zhongguo Zhong Yao Za Zhi. 2008 Feb;33(4):359-62.
To establish an effective way for rapid identification of Monascus strains based on DNA molecular marker.
A random amplified polymorphic DNA (RAPD) marker named F421 in genomic DNA of Monascus F strain was observed during a comparison of DNA fingerprints derived from 10 cultivated strains of Monascus. F421 was cloned and sequenced. Comparing the sequence of F421 (GenBank accession number EF063107) with other relative sequences in the GenBank databases, no distinct comparability was found. A pair of sequence characterized amplified region (SCAR) primers were designed based on the sequence of the cloned fragment and tested for the specific detection of Monascus F.
The results of polymerase chain reaction showed that only a 421bp segment of Monascus F strain was amplified compared with other 9 cultivated strains of Monascus. And the acquired SCAR marker of strain F could be used as a specific DNA fingerprint to identify Monascus strain F within one day.
SCAR molecular marker technology is an effective new way to identify Monascus strains more rapidly. And also is an assistant tool to identify Monascus strains more accurately when disagreements come out using traditional classification. It could be applied widely to the protection of germ plasm resources, classification and identification distinguishing false strains of pharmaceutical fungi.
建立一种基于DNA分子标记快速鉴定红曲霉菌株的有效方法。
在对10株红曲霉菌株的DNA指纹图谱进行比较时，在红曲霉菌F株基因组DNA中观察到一个名为F421的随机扩增多态性DNA（RAPD）标记。对F421进行克隆和测序。将F421的序列（GenBank登录号EF063107）与GenBank数据库中的其他相关序列进行比较，未发现明显的可比性。根据克隆片段的序列设计了一对序列特异性扩增区域（SCAR）引物，并对红曲霉菌F进行特异性检测。
聚合酶链反应结果显示，与其他9株红曲霉菌株相比，仅扩增出红曲霉菌F株的一个421bp片段。获得的菌株F的SCAR标记可作为特异性DNA指纹图谱，在一天内鉴定红曲霉菌株F。
SCAR分子标记技术是一种更快速鉴定红曲霉菌株的有效新方法。也是在传统分类出现分歧时更准确鉴定红曲霉菌株的辅助工具。它可广泛应用于种质资源保护、分类鉴定以及区分药用真菌的假菌株。
