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 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.
D010582:Bacteriophage lambda / Q000235:genetics; D002874:Chromosome Mapping; D002897:Chromosomes, Human, Pair 7; D003001:Cloning, Molecular; D003550:Cystic Fibrosis / Q000235:genetics*; D004247:DNA / Q000235:genetics*; D004586:Electrophoresis; D005819:Genetic Markers; D006801:Humans; D009693:Nucleic Acid Hybridization; D009857:Oncogenes
D005819:Genetic Markers; D004247:DNA