快速聚合酶链反应(PCR)-单链构象多态性(SSCP)筛选法检测钙调素核苷酸变异对黑曲霉种进行鉴定。

A Susca, G Stea, G Perrone
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引用次数: 20

摘要

单链构象多态性(SSCP)分析已广泛应用于遗传多样性研究,用于间接检测相同长度的扩增DNA片段中单个碱基的序列差异,是基因测序的一种替代方法。本研究采用SSCP分析法检测钙调蛋白基因约180 bp区域的序列变异,以鉴定Aspergillus section Nigri种。所描述的方法表明,基于荧光的毛细管电泳SSCP分析比直接测序更便宜、更快,并且适用于计算机辅助分析,可以区分属于黑曲霉组的曲霉种:A. aculeatus、A. carbonarius、A. ellipticus、A. foetidus、A. heteromorphus、A. ibericus、A. japonicus、A. niger和A. tubingensis。
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Rapid polymerase chain reaction (PCR)-single-stranded conformational polymorphism (SSCP) screening method for the identification of Aspergillus section Nigri species by the detection of calmodulin nucleotide variations.

Single-stranded conformational polymorphism (SSCP) analysis for genetic diversity studies has been widely applied to detect indirectly sequence differences up to a single base in amplified DNA fragments of the same length, representing an alternative to gene sequencing. In this study SSCP analysis was used to detect sequence variations contained in an about 180-bp region of the calmodulin gene in order to identify Aspergillus section Nigri species. The method described shows that fluorescence-based SSCP analysis by capillary electrophoresis is cheaper and faster than direct sequencing, and suitable for computer-assisted analyses allowing discrimination between the Aspergillus species belonging to the Nigri section: A. aculeatus, Aspergillus 'atypic uniseriate', A. brasiliensis, A. carbonarius, A. ellipticus, A. foetidus, A. heteromorphus, A. ibericus, A. japonicus, A. niger, and A. tubingensis.

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