Abstract |
Laser microdissection has been used to dissect material from the X-chromosome region involved in fragile- X-linked mental retardation. After dissection, single chromosome slices corresponding to this fragile site were subjected to DNA amplification using either a vector ligation method (to provide known anchor sequences) or primer oligonucleotides corresponding to the ubiquitous Alu sequences. Amplified material was then cloned or, alternately, used to screen a gridded cosmid library. Eight cosmid clones identified in this way were regionally mapped using a panel of hybrid cell lines and shown to originate from a narrow interval centered on the fragile X site. Two clones are included in the approximately 6-cM interval defined by probes RNI (DXS369, 5 cM proximal) and VK21 (DXS 296, 1-2 cM distal) and which includes the fragile site, and at least one clone contains sequences conserved across species suggestive of a gene. This method combines the focused approach of microdissection and the convenience of obtaining cosmid (rather than small-insert) clones; it may be useful for studies of other defined chromosomal regions.
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Authors | M Djabali, C Nguyen, I Biunno, B A Oostra, M G Mattei, J E Ikeda, B R Jordan |
Journal | Genomics
(Genomics)
Vol. 10
Issue 4
Pg. 1053-60
(Aug 1991)
ISSN: 0888-7543 [Print] United States |
PMID | 1916812
(Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Oligodeoxyribonucleotides
- DNA
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Topics |
- Animals
- Base Sequence
- Chromosome Banding
- Cloning, Molecular
(methods)
- Cosmids
- DNA
(genetics)
- Fragile X Syndrome
(genetics)
- Gene Library
- Genetic Vectors
- Humans
- Lasers
- Male
- Molecular Sequence Data
- Oligodeoxyribonucleotides
- Polymerase Chain Reaction
(methods)
- Restriction Mapping
- X Chromosome
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