A region-specific microdissection library for human chromosome 2p23--p21 and the analysis of an interstitial deletion of 2p21
| Autor: | J. Qi, F.T. Kao, M. Muenke, S. Tong, J. Yu |
|---|---|
| Rok vydání: | 1995 |
| Předmět: |
Positional cloning
Base pair Restriction Mapping CHO Cells Deoxyribonuclease HindIII Biology Hybrid Cells Genome law.invention Cell Line Embryonic and Fetal Development Gene mapping law Mesencephalon Cricetinae Holoprosencephaly Genetics Animals Humans Genomic library Cloning Molecular Molecular Biology Genetics (clinical) Microdissection Polymerase chain reaction Gene Library Dissection Chromosome Mapping Molecular biology Chromosomes Human Pair 2 Face Chromosome Deletion Low copy number |
| Zdroj: | Cytogenetics and cell genetics. 68(1-2) |
| ISSN: | 0301-0171 |
| Popis: | A region-specific library for human chromosome 2p23–p25 was constructed using microdissection and polymerase chain reaction (PCR)-mediated microcloning techniques. This library is large, comprising 300,000 recombinant microclones. The insert sizes range between 50–600 base pairs (bp) with a mean of 200 bp. About 50%–60% of the clones contain unique or very low copy number sequence inserts as determined by their weak or no hybridization to total human DNA. A subset of 48 microclones that did not hybridize to total human DNA after colony hybridization was analyzed, and 26 (54%) clones were shown to contain single-copy inserts and hybridize to human chromosome 2 DNAs, indicating that they are human chromosome 2 specific. The human genomic fragments identified by these clones after cleavage with HindIII have also been characterized. The single-copy microclones were used to analyze an interstitial deletion in the 2p23.3–p25.1 region — 46,XY, del(2) (pter→p25.1::p23.3→qter) — previously reported in a patient with severe growth and mental retardation and multiple anomalies. Of the 26 microclones analyzed, 14 clones were mapped to the deletion region. The availability of the 2p23–p25 region-specific library and the probes derived from the library should be valuable for fine structure physical mapping analysis and the cloning of disease-related genes localized to the region. These studies also demonstrate the efficiency with which useful probes can be quickly generated for genome studies and for positional cloning. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|