Orbach M J, Vollrath D, Davis R W, Yanofsky C
Department of Biological Sciences, Stanford University, California 94305-5020.
Mol Cell Biol. 1988 Apr;8(4):1469-73. doi: 10.1128/mcb.8.4.1469-1473.1988.
A molecular karyotype of Neurospora crassa was obtained by using an alternating-field gel electrophoresis system which employs contour-clamped homogeneous electric fields. The migration of all seven N. crassa chromosomal DNAs was defined, and five of the seven molecules were separated from one another. The estimated sizes of these molecules, based on their migration relative to Schizosaccharomyces pombe chromosomal DNA molecules, are 4 to 12.6 megabases. The seven linkage groups were correlated with specific chromosomal DNA bands by hybridizing transfers of contour-clamped homogeneous electric field gels with radioactive probes specific to each linkage group. The mobilities of minichromosomal DNAs generated from translocation strains were also examined. The methods used for preparation of chromosomal DNA molecules and the conditions for their separation should be applicable to other filamentous fungi.
利用采用轮廓夹钳均匀电场的交变电场凝胶电泳系统获得了粗糙脉孢菌的分子核型。确定了所有七条粗糙脉孢菌染色体DNA的迁移情况,并且将七条分子中的五条彼此分离开来。根据它们相对于粟酒裂殖酵母染色体DNA分子的迁移情况估算,这些分子的大小为4至12.6兆碱基。通过用每个连锁群特异的放射性探针与轮廓夹钳均匀电场凝胶的转移产物杂交,将七个连锁群与特定的染色体DNA条带关联起来。还检测了由易位菌株产生的微型染色体DNA的迁移率。制备染色体DNA分子所采用的方法及其分离条件应该适用于其他丝状真菌。