期刊名称:Proceedings of the National Academy of Sciences
印刷版ISSN:0027-8424
电子版ISSN:1091-6490
出版年度:1974
卷号:71
期号:7
页码:2823-2827
DOI:10.1073/pnas.71.7.2823
语种:English
出版社:The National Academy of Sciences of the United States of America
摘要:Eco R1 restriction endonuclease makes two cuts in each repeating unit of amplified ribosomal DNA (rDNA) from Xenopus laevis. The locations of these cuts have been established by comparison of the secondary structure of single-stranded Eco R1 fragments (as visualized in the electron microscope) with that of X. laevis rRNAs and of long strands of uncut rDNA. Of the two classes of fragments generated, the smaller one contains 90% of the 28S rRNA gene, has a duplex molecular weight of 3.0 x 106 and is homogeneous in size. The larger class of molecules contains 80% of the 18S rRNA gene and all of the nontranscribed spacer. These latter fragments are heterogeneous with molecular weights ranging from 4.0 to 5.9 x 106. The distribution of sizes within the large class of fragments varies among different preparations of rDNA, and heterogeneity is present in the DNA amplified from the rDNA of a single nucleolar organizer. The heterogeneity is located in the nontranscribed spacer region which is variable in length. This has been demonstrated by the formation of deletion loops in heteroduplexes made between larger fragments of different lengths.
关键词:Eco R1 restriction endonuclease ; gel electrophoresis ; electron microscopy ; secondary structure maps ; genes