Laser microdissection of the fragile X region: identification of cosmid clones and of conserved sequences in this region.
Genomics
; 10(4): 1053-60, 1991 Aug.
Article
em En
| MEDLINE
| ID: mdl-1916812
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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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Cromossomo X
/
Cosmídeos
/
Síndrome do Cromossomo X Frágil
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
/
Humans
/
Male
Idioma:
En
Ano de publicação:
1991
Tipo de documento:
Article