Summary
A male-specific 5.7 kb DNA(pKY-1) was cloned by screening human genomic libraries. Subsequent to structural characterization, a possibility of its application for prenatal diagnosis probe was tested. The results obtained were as follows: 1) The pKY-1 was specifically hybridized with human Y-chromosome specific repetitive 3.5 kb, but not with female DNA and 2.1 kb, known to be another human Y-specific repetitive-DNA. 2)HaeIII,EcoRI and their double digestion did not produce the Y-specific 3.5 kb fragments from pKY-1. 3) Although the digestion of pKY-1 withBamHI liberated 0.55 kb,ClaI clove remained 5.10 kb into 2.40 and 2.7 kb fragments, while their hybridizabilities with the 3.5 kb DNA were identically conserved.
These data suggest that 5.7 kb fragment (pKY-1) should be proposed as a member of Y-chromosome-specific repetitive DNA family in human genome. Thus, this cloned DNA will be not only a useful probe for human fetal sex-determination but also an effective tool for analysis of Y-chromosome with its structure and repetitive DNA.
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Abbreviations
- kb:
-
103 bases
- SSC:
-
0.15m sodium chloride-0.015m sodium citrate
- SDSa:
-
sodiumN-lauroylsarcosinate
- EDTA:
-
ethylenediaminetetraacetic acid sodium salts
References
Birnboim, H.C. and Doly, J. 1979. Rapid alkaline extraction procedure for screening recombinant plasmid DNA.Nucleic Acid Res. 7:1513–1523.
Cooke, H.J., Schmidtke, J. and Gosden, J.R. 1982. Characterization of human Y-chromosome repeated sequence and related sequences in higher primates.Chromosoma 87:491–502.
Gänshirt, D. and Pawlowitzki, I.H. 1984.HaeIII restriction of DNA from three cases with non fluorescent Y-chromosomes (45XO/46XYnf).Hum. Genet. 67:241–244.
Gosden, J.R., Gosden, C.M., Christie, S. and Morsman, J.M. 1984a. Rapid fetal sex determination in first trimester prenatal diagnosis by dot hybridization.Lancet I:540–541.
Gosden, J.R., Gosden, C.M., Christie, S., Cooke, H.J., Morsman, J.M. and Rodeck, C.H. 1984b. The use of cloned Y-chromosome-specific DNA probes for fetal sex-determination in first trimester prenatal diagnosis.Hum. Genet. 66:347–351.
Gross-Bellard, M., Oudet, P. and Chambon, P. 1973. Isolation of high-molecular-weight DNA from mammalian cells.Eur. J. Biochem. 36:32–38.
Kelly, R.B., Cozzarelli, N.R., Deustcher, M.P., Lehman, I.R. and Konberg, A. 1970. Enzymatic synthesis of deoxyribonucleic acid (labelling of DNA with32P by nick translation).J. Biol. Chem. 245:39–45.
Lau, Y.F., Huang, J.C., Dozy, A.M. and Kan, Y.W. 1984. A rapid screening test for antenatal sex determination.Lancet II:14–16.
Lau, Y.F. 1985. Detection of Y-specific repeat sequences in normal and variant human chromosomes usingin situ hybridization with biotinylated probes.Cytogenet Cell Genet. 39:184–187.
Maniatis, T., Fritsch, E.F., and Sambrook, J. 1982.Molecular Cloning—A Laboratory Manual. Cold Spring Harber Laboratory, Cold Spring Harbor, New York, pp. 164–165.
Nagai, K., Koike, S., Kominami, R., Muramatsu, M., Soma, H. and Yanagisawa, I. 1984. Cloning of human Y chromosome specific DNA. The 7th Meeting of the Molecular Biology Society of Japan (abstract): 19.
Nagai, K., Kominami, R. and Yanagisawa, I. 1986. The characterization of human Y-chromosome-specific cloned 5.7 kb DNA.Biol. Chem. Hoppe-Seyler,367, (Supple.):163.
Nakahori, Y. Mitani, K., Yamada, M. and Nakagome, Y. 1986. A human Y-chromosome specific repeated DNA family (DYZ1) consist of a tandem array of pentanucleotides.Nucleic Acids Res. 14:7569–7580.
Southern, E.M. 1975. Detection of specific sequences among DNA fragments separated by gel electrophoresis.J. Mol. Biol. 98:503–517.
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Nagai, K., Koike, S., Kominami, R. et al. The characterization of human male-specific 5.7 kb DNA(pKY-1) and its application to fetal sex-determination probe. Jap J Human Genet 34, 261–267 (1989). https://doi.org/10.1007/BF01929207
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DOI: https://doi.org/10.1007/BF01929207