论文部分内容阅读
对脉冲电泳研究中用做估测样本 DNA 分子量大小(即做为分子量标准)的 2 种酵母菌完整染色体 DNA 的不同制备方法进行了比较,同时利用脉冲电泳研究了几种植物病原真菌的分子核型,并探讨了植物病原真菌完整染色体 DNA 的简化制备方法。研究证实,制备啤酒酵母(Saccharomyces cerevisiae)染色体DNA 分子量标记的凝胶包埋破壁法易获得大量染色体 DNA,且用国产蜗牛酶(Snailase)可代替进口裂解酶(Driselase),SDS可代替十二烷基肌氨酸(N-Lau-roylsarcosine),降低了成本;改进的液氮冷冻研磨法与凝胶包埋破壁法具有相同的良好效果,得到了啤酒酵母全部16 条染色体 DNA。该方法的优点是制备程序简便、处理时间短、成本低,可完全取代凝胶包埋破壁法;凝胶包埋电泳法(即不破除细胞壁)效果不佳,不易获得完整染色体 DNA。首次利用液氮冷冻研磨法制备粟酒裂殖酵母(Schizosaccharomyces pom be)染色体DNA 分子量标记,得到全部 3 条完整染色体DNA,获得良好结果,为脉冲电泳研究中较大 DNA 分子量标记的快速简便制备奠定了基础。对几种植物病原真菌的分子核型进行了初步研究,获得了其基因组结构的基本数据。除串珠镰孢菌(Fusariumm oniliform e)和茄镰孢菌(F.solani)外,禾谷镰孢菌(F.gram inearum )、棉枯萎病菌(F.oxysporum f
The different preparation methods of the intact chromosomal DNA of two kinds of yeast used in the pulse electrophoresis research to estimate the molecular weight of the sample DNA (ie, as the molecular weight standard) were compared. Meanwhile, the molecular nuclei of several plant pathogenic fungi Type and discussed the simplified preparation method of the complete chromosomal DNA of plant pathogenic fungi. Studies have confirmed that the preparation of the molecular weight markers of chromosome DNA Saccharomyces cerevisiae gel-embedded broken method easy access to a large number of chromosomal DNA, and the use of domestic snail enzyme (Snailase) can replace imported lyase (Driselase), SDS can replace twelve N-Lau-roylsarcosine decreased the cost. The modified liquid nitrogen freezing method had the same good effect as the gel-encased method, and all the 16 chromosomal DNAs of S. cerevisiae were obtained. The method has the advantages of simple preparation process, short processing time and low cost, and completely replaces the gel-embedded wall breaking method; and the gel-embedded electrophoresis method (ie, not breaking the cell wall) is ineffective and the complete chromosomal DNA is not easily obtained. For the first time, molecular weight markers of Schizosaccharomyces pom be chromosome DNA were prepared by liquid nitrogen freezing and grinding method, and all three complete chromosomal DNAs were obtained. Good results were obtained, which provided a quick and easy preparation for larger DNA molecular weight markers in pulse electrophoresis The foundation. The molecular karyotypes of several plant pathogenic fungi were preliminary studied, and the basic data of their genome structure were obtained. In addition to Fusarium m oniliform e and F. solani, F. graminearum, F. oxysporum f.