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Nucleic Acids Research, 1983, Vol. 11, No. 20 6965-6983
© 1983

MOLECULAR BIOLOGY

Construction of a small Mus musculus repetitive DNA library: identification of a new satedllite sequence in Mus musculus

D.F. Pietras2, K.L. Bennett3, L.D. Siracusa1, M. Woodworth-Gutai, V.M. Chapman1, K.W. Gross1, C. Kane-Haas and N.D. Hastie4,*

Departments of Cell and Tumor Biology, Roswell Park Memorial Institute Buffalo, NY 14203, USA 1Molecular Biology, Roswell Park Memorial Institute Buffalo, NY 14203, USA

*To whom correspondence should be addressed

Received August 8, 1983. Accepted September 23, 1983.

We report the construction of a small library of recombinant plasmids containing Mus musculus repetitive DNA inserts. The repetitive cloned fraction was derived from denatured genomic DNA by reassociation to a Cot value at which repetitive, but not unique, sequences have reannealed followed by exhaustive S1 nuclease treatment to degrade single stranded DNA. Initial characterizations of this library by colony filter hybridizations have led to the identification of a prevously undetected M. musculus minor satellite as well as to clones containing M. musculus major satellite sequences. This new satellite is repeated 10–20 times less than the major satellite in the M. musculus genome. It has a repeat length of 130 nucleotides compared wih the M. musculus major satellite with a repeat length of 234 nucleotides. Sequence analysis of the minor satellite has shown that it has a 29 base pair region with extensive homology to one of the major satellite repeating subunits. We also show by in situ hybridization that this minor satellite sequence is located at the centromeres and possibly the arms of at least half the M musculus chromosomes. Sequences related to the minor satellite have been found in the DNA of a related Mus species, Mus spretus and may represent the major satellite of that species.

2Current address: Department of Radiation Biology and Biophysics, University of Rochester School of Medicine and Dentistry, Rochester, NY, USA

3Current address: Carnegie Institution of Washington, Department of Embryology, 115 West University Parkway, Baltimore, MD 21210, USA

4Current address: MRC Clinical and Population Cytogenetics Unit, Western General Hospital, Crewe Road, Edinburgh EH4 2XU, UK


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