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适用于多种蘑菇物种的反转录转座子微卫星扩增多态性菌株指纹图谱标记。

Retrotransposon microsatellite amplified polymorphism strain fingerprinting markers applicable to various mushroom species.

作者信息

Le Quy Vang, Won Hyo-Kyung, Lee Tae-Soo, Lee Chang-Yun, Lee Hyun-Sook, Ro Hyeon-Su

机构信息

Department of Microbiology and Research Institute of Life Sciences, Gyeongsang National University, Chinju 660-701, Korea.

出版信息

Mycobiology. 2008 Sep;36(3):161-6. doi: 10.4489/MYCO.2008.36.3.161. Epub 2008 Sep 30.

Abstract

The retrotransposon marY1 is a gypsy family retroelement, which is detected ubiquitously within the fungal taxonomic groups in which mushrooms are included. To utilize marY1 as a molecular marker for the DNA fingerprinting of mushrooms, oligonucleotides marY1-LTR-L and marY1-LTR-R were designed on the basis of highly conserved regions from the multiple sequence alignment of 30 marY1 sequences retrieved from a nucleotide sequence database. In accordance with Retrotransposon Microsatellite Amplified Polymorphism (REMAP) fingerprinting methodology, the two oligonucleotides were utilized together with the short sequence repeat primers UBC807 and UBC818 for polymerase chain reaction using templates from different mushroom genomic DNAs. Among the tested oligonucleotides, the marY1-LTR-L and UBC807 primer set yielded the greatest amount of abundance and variation in terms of DNA band numbers and patterns. This method was successfully applied to 10 mushroom species, and the primer set successfully discriminated between different commercial mushroom cultivars of the same strains of 14 Pleurotus ostreatus and 16 P. eryngii. REMAP reproducibility was superior to other popular DNA fingerprinting methodologies including the random amplified polymorphic DNA method.

摘要

逆转座子marY1是一种gypsy家族逆转元件,在包括蘑菇在内的真菌分类群中普遍存在。为了将marY1用作蘑菇DNA指纹识别的分子标记,基于从核苷酸序列数据库检索到的30个marY1序列的多序列比对中的高度保守区域,设计了寡核苷酸marY1-LTR-L和marY1-LTR-R。根据逆转座子微卫星扩增多态性(REMAP)指纹识别方法,将这两种寡核苷酸与短序列重复引物UBC807和UBC818一起用于聚合酶链反应,使用来自不同蘑菇基因组DNA的模板。在测试的寡核苷酸中,marY1-LTR-L和UBC807引物组在DNA条带数量和模式方面产生了最大量的丰度和变异。该方法成功应用于10种蘑菇物种,并且该引物组成功区分了14种平菇和16种刺芹侧耳相同菌株的不同商业蘑菇品种。REMAP的重现性优于其他流行的DNA指纹识别方法,包括随机扩增多态性DNA方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/365d/3755187/b88acf017eb8/mb-36-161-g001.jpg

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