Weising K, Beyermann B, Ramser J, Kahl G
Pflanzliche Molekularbiologie, Fachbereich Biologie, Johann Wolfgang Goethe-Universität, Frankfurt, Germany.
Electrophoresis. 1991 Feb-Mar;12(2-3):159-69. doi: 10.1002/elps.1150120211.
The existence of hypervariable DNA sequences in nuclear genomes, and the use of appropriate "fingerprinting" probes to detect them, has gained widespread scientific interest, and also led to multiple applications in diverse areas. Two years ago, the new technique of "DNA fingerprinting" was also introduced into the analysis and characterization of plant genomes, initially by using human or M13 minisatellites as probes. In the present article, we demonstrate the applicability for plant DNA fingerprinting of oligonucleotide probes specific for simple repetitive DNA sequences. We show that various levels of intra- and interspecific polymorphisms can be detected; the information to be gained depends on the optimal combination of probe and species. Variety-specific patterns were obtained in several cases. Some probes revealed variability between individuals. Somatic variability was not observed. Different DNA isolation and purification procedures were tested in order to introduce a fast and easy-to-perform isolation method suitable for a large variety of plant species. Nonradioactive fingerprinting was performed using digoxigenated oligonucleotides as probes. Banding patterns obtained with radioactive and digoxigenin-based labeling techniques proved to be of similar quality.
核基因组中高变DNA序列的存在,以及使用合适的“指纹”探针来检测它们,已引起广泛的科学关注,并在不同领域有了多种应用。两年前,“DNA指纹识别”新技术也被引入植物基因组的分析和特征描述中,最初是使用人类或M13微卫星作为探针。在本文中,我们证明了针对简单重复DNA序列的寡核苷酸探针在植物DNA指纹识别中的适用性。我们表明,可以检测到种内和种间不同水平的多态性;所获得的信息取决于探针和物种的最佳组合。在几种情况下获得了品种特异性模式。一些探针揭示了个体之间的变异性。未观察到体细胞变异性。测试了不同的DNA分离和纯化程序,以引入一种适用于多种植物物种的快速且易于操作的分离方法。使用地高辛标记的寡核苷酸作为探针进行非放射性指纹识别。用放射性和基于地高辛的标记技术获得的条带模式质量相似。