Construction of a General Human Chromosome Jumping Library, with Application to Cystic Fibrosis

In many genetic disorders, the responsible gene and its protein product are unknown. The technique known as ``reverse genetics,'' in which chromosomal map positions and genetically linked DNA markers are used to identify and clone such genes, is complicated by the fact that the molecular d...

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Bibliographic Details
Published in:Science (American Association for the Advancement of Science) 1987-02, Vol.235 (4792), p.1046-1049
Main Authors: Collins, Francis S., Drumm, Mitchell L., Cole, Jeffery L., Lockwood, Wendy K., George F. Vande Woude, Iannuzzi, Michael C.
Format: Article
Language:English
Subjects:
Bacteriophage lambda - genetics
Bacteriophages
Biological and medical sciences
Biotechnology
Chromosome Mapping
Chromosomes
Chromosomes, Human, Pair 7
Classical genetics, quantitative genetics, hybrids
Cloning, Molecular
Cystic fibrosis
Cystic Fibrosis - genetics
DNA
DNA - genetics
DNA probes
Electrophoresis
Fundamental and applied biological sciences. Psychology
Gels
gene libraries
Gene therapy
Genes
Genetic aspects
Genetic code
Genetic Markers
Genetics
Genetics of eukaryotes. Biological and molecular evolution
Genomes
Genomics
Health. Pharmaceutical industry
Human
Human chromosomes
Humans
Industrial applications and implications. Economical aspects
Libraries
man
Methods
Nucleic Acid Hybridization
Oncogenes
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Summary:In many genetic disorders, the responsible gene and its protein product are unknown. The technique known as ``reverse genetics,'' in which chromosomal map positions and genetically linked DNA markers are used to identify and clone such genes, is complicated by the fact that the molecular distances from the closest DNA markers to the gene itself are often too large to traverse by standard cloning techniques. To address this situation, a general human chromosome jumping library was constructed that allows the cloning of DNA sequences approximately 100 kilobases away from any starting point in genomic DNA. As an illustration of its usefulness, this library was searched for a jumping clone, starting at the met oncogene, which is a marker tightly linked to the cystic fibrosis gene that is located on human chromosome 7. Mapping of the new genomic fragment by pulsed field gel electrophoresis confirmed that it resides on chromosome 7 within 240 kilobases downstream of the met gene. The use of chromosome jumping should now be applicable to any genetic locus for which a closely linked DNA marker is available.
ISSN:0036-8075
1095-9203
DOI:10.1126/science.2950591