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以早熟白菜苔为实验材料,从其基因组DNA中分离出C0t-1 DNA并用生物素标记作探针,25S rDNA用地高辛标记作探针,对有丝分裂中期相染色体进行双色荧光原位杂交。每对染色体上均显示出了特定的C0t-1 DNA荧光原位杂交带型,5对染色体上显示出了25S rDNA荧光原位杂交带型。双色荧光原位杂交证实了C0t-1 DNA与25S rDNA二者具有一致的染色体位置特征,表明基于rDNA及C0t-1 DNA的荧光原位杂交核型分析技术,优于目前普遍采用的只基于rDNA的荧光原位杂交核型分析方法。结合C0t-1 DNA与25S rDNA的荧光原位杂交带型和传统的染色体的形态学标记分析方法及白菜已公布的基于rDNA分布的核型分析结果,创建了一个精确的白菜核型。
Taking precocious cabbage moss as experimental material, C0t-1 DNA was isolated from its genomic DNA and labeled with biotin. 25S rDNA was labeled with digoxigenin as a probe to detect mitochondrial metaphase chromosome by two-color fluorescence in situ hybridization. Each pair of chromosomes displayed a specific C0t-1 DNA fluorescence in situ hybridization pattern, and five pairs of chromosomes showed 25S rDNA fluorescence in situ hybridization patterns. Two-color fluorescence in situ hybridization confirmed that both C0t-1 DNA and 25S rDNA had the same chromosomal location characteristics, indicating that the fluorescence in situ hybridization karyotyping technology based on rDNA and C0t-1 DNA is superior to the commonly used method based on rDNA Fluorescence in situ hybridization karyotyping method. Combining the fluorescence in situ hybridization patterns of C0t-1 DNA and 25S rDNA with traditional chromosome morphological analysis and the published karyotype analysis results based on rDNA distribution in cabbage, an accurate karyotype was established.