Yao M C
Cell. 1981 Jun;24(3):765-74. doi: 10.1016/0092-8674(81)90102-1.
The chromosomal DNA sequence adjacent to one end of the single ribosomal RNA gene (rDNA) in the micronucleus of Tetrahymena has been isolated by cloning. Using this sequence as a hybridization probe the organization of the same sequence in the somatic macronucleus has been examined. The restriction enzyme digestion maps of this sequence in the two nuclei are very different. Detailed mapping studies suggest that a chromosome break has occurred near the junction between the rDNA and the neighboring sequence during the formation of the macronucleus. As a result the flanking sequence is located near a free chromosome end in the macronucleus. The existence of such a linear DNA end has also been shown by digestion with the exonuclease Bal 31. In addition to the breakage, some sequences at this junction are found to be eliminated from the macronucleus. This observation has been interpreted in relation to the mechanism of rDNA amplification, which in Tetrahymena generates extrachromosomal rDNA molecules during macronucleus development.
通过克隆已分离出四膜虫微核中与单个核糖体RNA基因(rDNA)一端相邻的染色体DNA序列。以该序列作为杂交探针,研究了其在体细胞大核中的组织情况。该序列在两个核中的限制性内切酶消化图谱差异很大。详细的图谱研究表明,在大核形成过程中,rDNA与相邻序列的连接处附近发生了染色体断裂。结果,侧翼序列位于大核中一个游离染色体末端附近。用核酸外切酶Bal 31消化也证明了这种线性DNA末端的存在。除了断裂外,还发现该连接处的一些序列在大核中被消除。这一观察结果已根据rDNA扩增机制进行了解释,在四膜虫中,rDNA扩增在大核发育过程中产生染色体外rDNA分子。