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 language | English |
|---|---|
| Pages (from-to) | 1053-1060 |
| Number of pages | 8 |
| Journal | Genomics |
| Volume | 10 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - 1991 |
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ASJC Scopus subject areas
- Genetics
Cite this
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 journal › Article
}
TY - JOUR
T1 - Laser microdissection of the fragile X region
T2 - Identification of cosmid clones and of conserved sequences in this region
AU - Djabali, Malek
AU - Nguyen, Catherine
AU - Biunno, Ida
AU - Oostra, B. A.
AU - Mattei, Marie Geneviève
AU - Ikeda, Joh E.
AU - Jordan, Bertrand R.
PY - 1991
Y1 - 1991
N2 - 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.
AB - 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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UR - http://www.scopus.com/inward/citedby.url?scp=0025737849&partnerID=8YFLogxK
U2 - 10.1016/0888-7543(91)90198-N
DO - 10.1016/0888-7543(91)90198-N
M3 - Article
C2 - 1916812
AN - SCOPUS:0025737849
VL - 10
SP - 1053
EP - 1060
JO - Genomics
JF - Genomics
SN - 0888-7543
IS - 4
ER -