Dong F, Miller J T, Jackson S A, Wang G L, Ronald P C, Jiang J
Department of Horticulture, University of Wisconsin, Madison, WI 53706, USA.
Proc Natl Acad Sci U S A. 1998 Jul 7;95(14):8135-40. doi: 10.1073/pnas.95.14.8135.
Rice bacterial artificial chromosome clones containing centromeric DNA were isolated by using a DNA sequence (pSau3A9) that is present in the centromeres of Gramineae species. Seven distinct repetitive DNA elements were isolated from a 75-kilobase rice bacterial artificial chromosome clone. All seven DNA elements are present in every rice centromere as demonstrated by fluorescence in situ hybridization. Six of the elements are middle repetitive, and their copy numbers range from approximately 50 to approximately 300 in the rice genome. Five of these six middle repetitive DNA elements are present in all of the Gramineae species, and the other element is detected only in species within the Bambusoideae subfamily of Gramineae. All six middle repetitive DNA elements are dispersed in the centromeric regions. The seventh element, the RCS2 family, is a tandem repeat of a 168-bp sequence that is represented approximately 6,000 times in the rice genome and is detected only in Oryza species. Fiber-fluorescence in situ hybridization analysis revealed that the RCS2 family is organized into long uninterrupted arrays and resembles previously reported tandem repeats located in the centromeres of human and Arabidopsis thaliana chromosomes. We characterized a large DNA fragment derived from a plant centromere and demonstrated that rice centromeres consist of complex DNA, including both highly and middle repetitive DNA sequences.
利用一种存在于禾本科物种着丝粒中的DNA序列(pSau3A9),分离出了含有着丝粒DNA的水稻细菌人工染色体克隆。从一个75千碱基对的水稻细菌人工染色体克隆中分离出了7种不同的重复DNA元件。荧光原位杂交表明,所有7种DNA元件都存在于每个水稻着丝粒中。其中6种元件是中度重复的,它们在水稻基因组中的拷贝数大约在50到300之间。这6种中度重复DNA元件中的5种存在于所有禾本科物种中,另一种仅在禾本科竹亚科的物种中被检测到。所有6种中度重复DNA元件都分散在着丝粒区域。第7种元件,即RCS2家族,是一个168碱基对序列的串联重复,在水稻基因组中大约有6000个拷贝,并且仅在稻属物种中被检测到。纤维荧光原位杂交分析表明,RCS2家族组织成长的不间断阵列,类似于先前报道的位于人类和拟南芥染色体着丝粒中的串联重复。我们对一个来自植物着丝粒的大DNA片段进行了表征,并证明水稻着丝粒由复杂的DNA组成,包括高度重复和中度重复的DNA序列。