在植物粗线期染色体和DNA纤维上的荧光原位杂交技术  

Fluorescence in situ Hybridization to PachyteneChromosomes and Extended DNA Fibres in Plants

作　　者：钟筱波 PaulFFransz JannieWennekes AbvanKammen JHansdeJong PimZabel 

机构地区：[1]荷兰瓦赫宁根农业大学分子生物学系 [2]荷兰瓦赫宁根农业大学遗传系

出　　处：《Acta Genetica Sinica》1998年第2期142-149,共8页

基　　金：荷兰国家自然科学基金会资助;欧洲共同体的资助

摘　　要：介绍了两种荧光原位杂交技术的详细实验步骤。第一种技术是在减数分裂粗线期染色体上的荧光原位杂交，包括从花粉母细胞中制备粗线期染色体和在这种染色体上定位ＤＮＡ序列，其分辨率水平能够达到１００ｋｂ。第二种技术是从植物细胞核中制备ＤＮＡ纤维，并在上面进行原位杂交，能够直接分析ＤＮＡ序列的分子排列关系，其分辨率水平能达到几个ｋｂ。为了说明这两种原位杂交技术在研究基因组和染色体结构、构建高分辨率的ＤＮＡ物理图谱上的能力，将展示用该技术直接分析番茄染色体端粒重复序列和端粒联接重复序列的染色体定位和ＤＮＡ分子排列。? This article describes two comprehensive fluorescence in situ hybridization (FISH) protocols for both pachytene chromosomes and extended DNA fibres in plants. The first technique involves FISH to spread pachytene chromosomes which are made from pollen mother cells, and proves to be an excellent method for assigning DNA sequences to chromosome regions at a resolution of up to a few hundred kilobases. The second technique produces an even higher FISH resolution on extended DNA fibres, prepared from interphase nuclei and used as hybridising component. This technique permits the delineation and ordering of contiguous sequences at strong enhancement of physical map resolution to values of a few kilobases. The power of both methods simultaneously applied for molecular cytogenetic analysis of genome organization and physical mapping was demonstrated with the combination of the telomeric repeat and the tomato specific telomereassociated repeat TGR1 as example.

关 键 词：荧光原位杂交 粗线期 染色体 DNA纤维 番茄 

分 类 号：Q943[生物学—植物学] Q78