A procedure for isolating transfer ribonucleic acid from human placenta
1970; Elsevier BV; Volume: 199; Issue: 2 Linguagem: Inglês
10.1016/0005-2787(70)90097-3
ISSN1879-3002
Autores Tópico(s)Molecular Biology Techniques and Applications
ResumoThe chapter discusses the fragmentation of ribosomes. It is one of the methods of investigating ribosomal structure obtained after partial cleavage of the ribosomal RNA. Fragments are analyzed to identify their proteins and nucleic acid sequences, whose presence in the same fragment indicates that they are located in the same restricted region of the intact ribosome. A considerable amount of information of this sort is published for both ribosomal subunits of Escherichia coli. In conjunction with information obtained with other methods, this information is contributing to the on-going process of constructing a three-dimensional map of the ribosomal components. The approach is based on the assumption that the conformation and stability of ribosomes and ribosomal fragments are the net result of the various mutual interactions among RNA, proteins, and the ions of the buffer. A single buffer is employed throughout the entire process, including all preparative and analytical procedures. After cleavage of the RNA, the ribosome is dissociated into fragments by heat rather than by changing the medium. When these precautions were observed, a fragmentation pattern was obtained that could be reproduced routinely and whose various fractions were stable and maintained their sedimentation and electrophoretic properties throughout the several procedures used, making it possible to purify a fragment to homogeneity.
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