title: Isolation of region-specific and polymorphic markers from chromosome 17 by restricted Alu polymerase chain reaction.
abstract: We demonstrate that the digestion of template DNAs with restriction endonucleases prior to Alu polymerase chain reaction ("restricted Alu-PCR") reduces the complexity of the Alu-primed amplification patterns of human DNA in somatic cell hybrids and allows a direct informative comparison of these patterns. A comparison of restricted Alu-PCR patterns of a monochromosomal hybrid retaining a human chromosome 17 (MH22-6) and a hybrid retaining a human chromosome 17 deleted for band p11.2 (DH110-D1) revealed four Alu-PCR products that were present in the former but absent in the latter hybrid. Hybridization of these fragments to the total Alu-PCR amplification products of the two hybrids confirmed their absence in DH110-D1 amplification products. Hybridization to a panel of somatic cell hybrids indicated that two of these fragments were deleted in the hybrid DH110-D1 and mapped to 17p11.2, as expected. However, two additional fragments were not deleted in the hybrid DH110-D1 and mapped to other regions of chromosome 17. An insertion-deletion polymorphism was associated with one of the latter fragments, which may be the mechanism for the lack of its amplification in the hybrid DH110-D1. Restricted Alu-PCR should enhance the applications of Alu-PCR and provides a new method for the identification of chromosome-specific polymorphic markers.
authors.affiliation: Institute for Molecular Genetics, Baylor College of Medicine, Houston, Texas 77030.
authors.forename: V
authors.initials: V
authors.lastname: Guzzetta
authors.forename: R
authors.initials: R
authors.lastname: Montes de Oca-Luna
authors.forename: J R
authors.initials: JR
authors.lastname: Lupski
authors.forename: P I
authors.initials: PI
authors.lastname: Patel
mesh_terms: D000818:Animals; D002460:Cell Line; D002886:Chromosomes, Human, Pair 17; D005819:Genetic Markers; D006801:Humans; D006822:Hybrid Cells; D051379:Mice; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012091:Repetitive Sequences, Nucleic Acid
chemical_list: D005819:Genetic Markers