Isolation and chromosomal localization of unique DNA sequences from a human genomic library
| Autor: | Martha Liao Law, Jeffrey N. Davidson, Judith A. Hartz, Fa-Ten Kao |
|---|---|
| Rok vydání: | 1982 |
| Předmět: |
Genetics
Multidisciplinary Base Sequence biology DNA Recombinant EcoRI Chromosome Mapping Nucleic Acid Hybridization Hybrid Cells Coliphages Molecular biology DNA sequencing Nucleic acid thermodynamics Restriction map Cricetinae Escherichia coli biology.protein Animals Humans Genomic library Human genome Cloning Molecular Chromosome 22 Research Article Southern blot |
| Zdroj: | Proceedings of the National Academy of Sciences. 79:865-869 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.79.3.865 |
| Popis: | Recombinant bacteriophage lambda from a human genomic library were screened to indentify human DNA inserts having only unique sequences. Unique human inserts were found in about 1% of the phage screened. One recombinant phage, P3-2, was studied in detail. It contains a human insert of 14.7 kilobases with four internal EcoRI cleavage sites. A restriction map was constructed for EcoRI and BamHI sites. Hybridization of the 32P-labeled P3-2 probe to a Southern blot of EcoRI-digested total human DNA yielded distinct bands at positions corresponding to the human insert fragments contained in P3-2. By using a series of human-Chinese hamster somatic cell hybrids containing unique combinations of human chromosomes, the human DNA segment in phage P3-2 was assigned to human chromosome 22 by blot hybridization and synteny analysis. In addition, another human DNA segment, 11.4 kilobases, in phage P3-10 was assigned to human chromosome 10 by similar procedures. With this approach, more unique DNA sequences can be isolated, assigned to specific human chromosomes, and used as genetic markers for gene mapping and linkage, polymorphism, and other genetic studies in the human genome. |
| Databáze: | OpenAIRE |
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