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引用次数: 0
摘要
利用针对特定类型微生物的PCR分析很容易完成啤酒腐败微生物的鉴定,但利用培养独立的分子方法对野生酵母和酿造酵母进行一般分类仍然具有挑战性。本研究中提出的方法利用遗传指纹匹配来确定未知酵母菌分离物是否与已编目的酿造酵母菌菌株的指纹匹配。利用酵母转座子长末端重复序列(delta element)的PCR扩增,获得了delta Next Generation Sequencing (NGS)指纹图谱。利用开源软件对三角间PCR扩增子NGS产生的reads进行处理,生成识别不同酵母菌株的指纹图谱。三角洲间NGS指纹图谱包括可记录、比较和用作酵母菌株鉴定的固定参考的DNA序列。三角间NGS指纹图谱具有可重复性,能够区分酿酒酵母和野生酵母菌株。实验酵母污染证明了该方法在鉴别内部酿造酵母交叉污染和外来野生酵母污染方面的实用性。
Fingerprinting Saccharomyces cerevisiae Strains Using Next Generation Sequencing of PCR Amplicons Generated from Delta Elements
Abstract Identifying beer spoilage microbes is readily accomplished using PCR analyses targeting specific types of microbes, but the general classification of wild yeast versus brewing yeast using cultivation independent molecular methods has remained challenging. The approach presented in this study utilized genetic fingerprint matching to determine if an unknown yeast isolate matches the fingerprint of catalogued brewing yeast strains. Interdelta Next Generation Sequencing (NGS) fingerprints were produced using PCR amplification of delta elements, also known as long terminal repeat sequences of transposons of yeast. Fingerprints identifying different yeast strains were generated by processing reads produced by NGS of the interdelta PCR amplicons using open source software. The interdelta NGS fingerprint comprises DNA sequences that can be recorded, compared, and utilized as a fixed reference for yeast strain identification. Interdelta NGS fingerprints were shown to be reproducible and capable of distinguishing between strains of brewing yeast and wild yeast. Experimental yeast contamination demonstrated the utility of the approach for identifying in-house brewing yeast cross contamination versus foreign wild yeast contamination.
期刊介绍:
The Journal of the American Society of Brewing Chemists publishes scientific papers, review articles, and technical reports pertaining to the chemistry, microbiology, and technology of brewing and distilling, as well as the analytical techniques used in the malting, brewing, and distilling industries.