Specific cloning of DNA fragments absent from the DNA of a male patient with an X chromosome deletion - PubMed (original) (raw)

Specific cloning of DNA fragments absent from the DNA of a male patient with an X chromosome deletion

L M Kunkel et al. Proc Natl Acad Sci U S A. 1985 Jul.

Abstract

A method that allows the specific cloning of DNA fragments absent from patients homozygous or hemizygous for chromosomal deletions is described. The method involves phenol-accelerated competitive DNA reassociation and subsequent molecular cloning of appropriately reassociated molecules. The deletion DNA sample utilized in the competition was isolated from a patient with a minute interstitial deletion in the short arm of the X chromosome. Sheared DNA isolated from a male child, who was diagnosed as having Duchenne muscular dystrophy, chronic granulomatous disease, and retinitis pigmentosa, was combined in a 200-fold excess with Mbo I-cleaved DNA isolated from a 49, XXXXY human lymphoid cell line, and the mixture was subjected to a phenol-enhanced reassociation technique. Analysis of 81 unique segments derived from cloned reassociated DNA molecules has led to the identification of 4 (5%) human DNA fragments that are absent from the male patient's DNA. The 4 clones were localized, on the basis of hybridization with restriction nuclease-digested genomic DNA from a panel of human and human-rodent hybrid cell lines, into three regions surrounding band 21 of the short arm of the normal human X chromosome. These clones are potential linkage markers for the diseases affecting this boy. Each clone, as well as others obtainable by this approach, may also serve as a starting point in the eventual cloning of these three X-linked-disease loci. Extension of this approach to other loci, including human tumors potentially homozygous for small deletions, should also be possible.

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References

1. Science. 1968 Aug 9;161(3841):529-40 - PubMed
1. Am J Hum Genet. 1985 Mar;37(2):250-67 - PubMed
1. Science. 1976 Mar 19;191(4232):1189-90 - PubMed
1. Biochemistry. 1977 Nov 29;16(24):5329-41 - PubMed
1. Am J Med Genet. 1979;3(1):43-58 - PubMed

Specific cloning of DNA fragments absent from the DNA of a male patient with an X chromosome deletion - PubMed (original) (raw)