Kempken F, Walz M
Lehrstuhl fur Allgemeine Botanik, Ruhr-Universitat, Bochum, D-44780, Germany
Fungal Genet Biol. 1996 Mar;20(1):89-92. doi: 10.1006/fgbi.1996.0015.
A method for the fast and reliable restriction analysis of intact fungal chromosomes is presented. For this purpose rare-cutting restriction enzymes of pro- and eukaryotic origin were used. Restriction fragments were separated on pulsed-field gels. Complete inactivation of proteinases remaining from chromosome preparations was crucial in this attempt. Rare-cutting restriction enzymes were employed to demonstrate the applicability of this procedure to the analysis of complex fungal genomes. This includes the localization of genes on discrete chromosomal fragments,the identification of telomeric restriction fragments, and the analysis of fungal transformants.
本文介绍了一种用于完整真菌染色体快速可靠限制性分析的方法。为此,使用了原核和真核来源的稀有切割限制性内切酶。限制性片段在脉冲场凝胶上进行分离。在这一尝试中,使染色体制备过程中残留的蛋白酶完全失活至关重要。使用稀有切割限制性内切酶来证明该方法在复杂真菌基因组分析中的适用性。这包括在离散染色体片段上定位基因、鉴定端粒限制性片段以及分析真菌转化体。