The use of standard and relaxed hybridization conditions to detect two classes of sequences related to type-D retroviruses in the DNAs of primates.

Using standard hybridization conditions (68 degrees C and 0.4 M sodium phosphate) and assaying for RNAase-resistant RNA . DNA duplexes in the presence of 2 X SSC (1 X SSC is 0.15 M NaCl and 0.015 M sodium citrate), sequences representing approx. 20% of the Mason-Pfizer virus (MPV) genome have previously been shown to be endogenous in DNAs of all Old World monkeys examined (Drohan, W., Colcher, D., Schochetman, G. and Schlom, J. (1977) J. Virol. 23, 36--43). We now report that titration of both the temperature at which hybridizations are carried out and the Na+ concentration at which 125I-labeled MPV RNA . DNA hybrids are scored, reveals a second class of sequences related to the MPV genome in the DNAs of primates. These MPV-related sequences, which are similar to an additional 40% of the MPV genome, are detected when the temperature of hybridization is reduced to 54 degrees C and when the resulting 125I-labeled RNA . DNA duplexes are scored for RNAase resistance in 8 X SSC. These sequences are found in the DNAs of all Old World monkeys examined, and the Tm values of the hybrid duplexes are approx. 6--7 degrees C lower than those of the hybrid duplexes formed using standard conditions. These studies further demonstrate the wide distribution of type-D retrovirus sequences in primates.