Affiliation: | a The University of Texas System Cancer Center M.D. Anderson Hospital and Tumor Institute, Department of Pathology, Section of Experimental Pathology, Houston, TX 77030, U.S.A. * Department of Cell Biology, Baylor College of Medicine, Houston, TX 77030, U.S.A. ** Department of Physics, The University of Texas System Cancer Center M.D. Anderson Hospital and Tumor Institute, Houston, TX 77030, U.S.A. |
Abstract: | The sequence of the 92 and 93 bp long, highly repetitive DNA fragments, isolated from EcoRI digested rat liver DNA, were determined. These fragments, designated 92 and 93, are found in equal abundance, 6.5 × 105 copies per haploid genome. J92 and J93 can be distinguished by their differential sensitivity to cleavage by HaeIII and HindIII, respectively, which cut the fragments at 75 and 57 bp from their mutually homologous 5'-ends. J92 and J93 are 38% and 35.4% G + C, respectively, and contain a disproportionate number of triplets complementary to stop codons in all reading frames. Three methylated sites were found in J92 while none could be detected in J93. The sequences around the m5C sites were 5'-Py-Py-m5C-G-Pu-Pu, except for one case where the second Py was replaced by an A. This site appeared to be hemimethylated. When J92 and J93 are placed in register from their mutually homologous 5'-ends, homology is 73% for the first 30 bp region and 63.5% for the total molecule. Thermal melting studies indicate sequence heterogeneity within J92 and J93 from substantial internal base mismatches. The sequences derived are therefore composite averages for the whole molecules. The Cot/1/2 for the sequence was measured spectrophotometrically to be 2 × 10−2 M/s on a DNA phosphorus basis and 2.15 × 10−4 M/s on a mole fragment basis. |