Purification and Some Characteristics of 5S DNA from Xenopus laevis

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RESUMO

DNA containing the multiple genes for 5S RNA has been isolated from the genome of Xenopus laevis. Whereas 5S RNA is about 57% G + C, the 5S DNA has a base composition of about 33-35% GC and consists of two alternating regions that differ in base composition by at least 20% GC. A denaturation map of 5S DNA analyzed by electron microscopy demonstrates that the repeating pattern is regular and each repeating unit has a mass of about 500,000 daltons. If one gene for 5S RNA (84,000 daltons native) were present in each repeat, it should comprise about 16.8% of 5S DNA. This arrangement is confirmed, since 6.8% of pure 5S DNA (13.6% of its base pairs) hybridized with 5S RNA. The remaining 83% of each repeating unit is considered to be “spacer” DNA. The 5S RNA hybridizes with about 0.05% of the bulk DNA of X. laevis, so that 5S DNA comprises about 0.7% of the total nuclear DNA. This is equivalent to about 24,000 repeating units for each haploid complement of DNA. These repeats are highly clustered; as many as 86 have been visualized along a single DNA molecule.

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