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首页> 外文期刊>Applied Microbiology and Biotechnology >Dereplication for biotechnology screening: PyMS analysis and PCR-RFLP-SSCP (PRS) profiling of 16S rRNA genes of marine and terrestrial actinomycetes
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Dereplication for biotechnology screening: PyMS analysis and PCR-RFLP-SSCP (PRS) profiling of 16S rRNA genes of marine and terrestrial actinomycetes

机译：用于生物技术筛选的重复复制：海洋和陆地放线菌的16S rRNA基因的PyMS分析和PCR-RFLP-SSCP（PRS）分析

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摘要

The search for exploitable biology is a major task for biotechnology-based industries. In this context, discrimination between previously tested or recovered micro-organisms (dereplication) is imperative, in order to reduce screening costs by sorting large collections of isolates, which are then subjected to further detailed evaluation. Pyrolysis mass spectrometry (PyMS) is a whole-cell fingerprinting technique that enables the rapid and reproducible sorting of micro-organisms, uses small samples and has the advantage of being fully automated. In this study, we compare chemometric fingerprinting with a ribotyping fingerprinting method, in order to investigate the extent to which pyrogroups formed by PyMS analysis relate to genetic diversity, using polymerase chain reaction-restriction fragment length polymorphism-single-strand conformational polymorphism (FIRS). A mixture of environmental strains of mycolic acid containing actinomycetes was used to mimic the selection of colonies from primary isolation plates. The congruence found between the clusters defined by the chemometric and molecular fingerprinting techniques was very high and demonstrated the effectiveness of PyMS as a rapid sorting and dereplicating procedure for putatively novel strains, criteria that are critical for biotechnological screens. Moreover, PyMS analysis revealed significant variation within pyrogroups that contained strains with the same genotypic (PRS) characteristics, thus emphasising its discriminatory capacity at the infraspecies level. [References: 30]
机译：寻找可利用的生物学是基于生物技术的产业的主要任务。在这种情况下，必须对以前测试过的微生物或回收的微生物（重复）进行区分，以便通过对大量分离株进行分选来降低筛选成本，然后对其进行进一步的详细评估。热解质谱法（PyMS）是一种全细胞指纹技术，可对微生物进行快速且可重复的分类，使用少量样品，并具有全自动的优势。在这项研究中，我们将化学计量学指纹图谱与核糖分型指纹图谱方法进行比较，以研究利用聚合酶链反应-限制性片段长度多态性-单链构象多态性（FIRS）通过PyMS分析形成的热基团与遗传多样性的关联程度。 。使用含有放线菌的霉菌酸的环境菌株的混合物来模拟从初级分离板中选择菌落。化学计量学和分子指纹技术定义的簇之间的一致性非常高，并证明了PyMS作为快速筛选和重复进行推定程序的有效性，该菌株对生物技术筛选至关重要。此外，PyMS分析显示，在含有相同基因型（PRS）特征的菌株的热基团中存在显着差异，从而强调了其在亚种水平上的区分能力。 [参考：30]

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