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J Virol. 1982 July; 43(1): 127-135

Most of the Murine Leukemia Virus Sequences in the DNA of NIH/Swiss Mice Consist of Two Closely Related Proviruses, Each Repeated Several Times

David L. Steffen¹, Richard Mural², Deborah Cowing^3,, Jane Mielcarz¹, Janet Young² and Richard Roblin²

¹ The Worcester Foundation for Experimental Biology, Shrewsbury, Massachusetts 01545
² Frederick Cancer Research Center, Frederick, Maryland 21701
³ Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139

ABSTRACT

The structure of the endogenous murine leukemia virus (MuLV) sequences of NIH/Swiss mice was analyzed by restriction endonuclease digestion, gel electrophoresis, and hybridization to an MuLV nucleic acid probe. Digestion of mouse DNA with certain restriction endonucleases revealed two classes of fragments. A large number of fragments (about 30) were present at a relatively low concentration, indicating that each derived from a sequence present once in the mouse genome. A smaller number of fragments (one to five) were present at a much higher concentration and must have resulted from sequences present multiple times in the mouse genome. These results indicated that the endogenous MuLV sequences represent a family of dispersed repetitive sequences. Hybridization of these same digested mouse DNAs to nucleic acid probes representing different portions of the MuLV genome allowed construction of a map of the sites where restriction endonucleases cleave the endogenous MuLV sequences. Several independent recombinant DNA clones of endogenous MuLV sequences have been isolated from C3H mice (Roblin et al., J. Virol. 43:113-126, 1982). Analysis of these sequences shows that they have the structure of MuLV proviruses. The sites at which restriction endonucleases cleave within these proviruses appeared to be similar or identical to the sites at which these nucleases cleaved within the MuLV sequences of NIH/Swiss mice. This identity was confirmed by parallel electrophoresis. We conclude that the apparently complex pattern of endogenous MuLV sequences of NIH/Swiss mice consists largely of only two kinds of provirus, each repeated multiple times at dispersed sites in the mouse genome.

Present address: Department of Molecular Biology, University of Wisconsin, Madison, WI 53703.