APPLICATION OF THE RANDOM AMPLIFIED POLYMORPHIC DNA USING THE POLYMERASE CHAIN-REACTION FOR EFFICIENT ELIMINATION OF DUPLICATE STRAINS IN MICROBIAL SCREENING .1. FUNGI
APPLICATION OF THE RANDOM AMPLIFIED POLYMORPHIC DNA USING THE POLYMERASE CHAIN-REACTION FOR EFFICIENT ELIMINATION OF DUPLICATE STRAINS IN MICROBIAL SCREENING .1. FUNGI
复制标题
DOI：
10.7164/antibiotics.47.173
复制
发表时间：
1994-02-01
影响因子：
3.3
通讯作者：
OKUDA, T
中科院分区：
文献类型：
--
作者：
FUJIMORI, F;OKUDA, T
为了在微生物筛选中有效地选择真菌菌株，我们应用了聚合酶链反应（PCR）的随机扩增多态性DNA（RAPD）方法。为了评价该系统，采用木霉属，因为其物种难以彼此区分。我们选择了合适的寡核苷酸十聚体R28（5 '-ATGGATCCGC），确定了PCR的最佳循环次数为30次，简化了模板制备方法，确定了最佳的模板浓度和Taq DNA聚合酶浓度。然后，我们检查了74个密切相关的木霉菌株。比较PCR产物的电泳带型。根据统计分析和系统发育分析（PAUP），结果与这些菌株的形态，生理和生态数据相一致。因此，RAPD技术是一种简便、有效、可靠的真菌菌株筛选方法。
For efficient fungal strain selection in microbial screening, we applied the random amplified polymorphic DNA (RAPD) method using the polymerase chain reaction (PCR). In order to evaluate this system, the genus Trichoderma was employed, because its species are difficult to distinguish from each other. We selected an appropriate oligonucleotide decamer, R28 (5'-ATGGATCCGC), determined the optimal cycles of PCR as 30 cycles, simplified the template preparation method, and determined optimal concentrations of the template and Taq DNA polymerase. We then examined 74 closely related strains of Trichoderma. The electrophoretic band patterns of the PCR products were compared. According to the statistical analysis with the phylogenetic analysis using parsimony (PAUP), the results were consistent with the morphological, physiological and ecological data on these strains. Therefore, we conclude that RAPD is a simple, efficient and reliable method for the selection of fungal strains employed in screening.