Laser microdissection of the fragile X region: Identification of cosmid clones and of conserved sequences in this region

Malek Djabali, Catherine Nguyen, Ida Biunno, B. A. Oostra, Marie Geneviève Mattei, Joh E. Ikeda, Bertrand R. Jordan

Research output: Contribution to journalArticle

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.

Original languageEnglish
Pages (from-to)1053-1060
Number of pages8
JournalGenomics
Volume10
Issue number4
DOIs
Publication statusPublished - 1991

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Microdissection
Cosmids
Conserved Sequence
Lasers
Clone Cells
X-Linked Mental Retardation
DNA Primers
Hybrid Cells
X Chromosome
Ligation
Dissection
Chromosomes
Cell Line
DNA
Genes

ASJC Scopus subject areas

  • Genetics

Cite this

Djabali, M., Nguyen, C., Biunno, I., Oostra, B. A., Mattei, M. G., Ikeda, J. E., & Jordan, B. R. (1991). Laser microdissection of the fragile X region: Identification of cosmid clones and of conserved sequences in this region. Genomics, 10(4), 1053-1060. https://doi.org/10.1016/0888-7543(91)90198-N

Laser microdissection of the fragile X region : Identification of cosmid clones and of conserved sequences in this region. / Djabali, Malek; Nguyen, Catherine; Biunno, Ida; Oostra, B. A.; Mattei, Marie Geneviève; Ikeda, Joh E.; Jordan, Bertrand R.

In: Genomics, Vol. 10, No. 4, 1991, p. 1053-1060.

Research output: Contribution to journalArticle

Djabali, M, Nguyen, C, Biunno, I, Oostra, BA, Mattei, MG, Ikeda, JE & Jordan, BR 1991, 'Laser microdissection of the fragile X region: Identification of cosmid clones and of conserved sequences in this region', Genomics, vol. 10, no. 4, pp. 1053-1060. https://doi.org/10.1016/0888-7543(91)90198-N
Djabali, Malek ; Nguyen, Catherine ; Biunno, Ida ; Oostra, B. A. ; Mattei, Marie Geneviève ; Ikeda, Joh E. ; Jordan, Bertrand R. / Laser microdissection of the fragile X region : Identification of cosmid clones and of conserved sequences in this region. In: Genomics. 1991 ; Vol. 10, No. 4. pp. 1053-1060.
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