title: Locus assignment of human alpha-globin structural mutants by selective enzymatic amplification of alpha 1 and alpha 2-globin cDNAs.
abstract: We have used the powerful methodology of DNA enzymatic amplification in order to assign human alpha-globin structural mutants to one of the two highly homologous alpha-globin genes. Selectively amplified alpha 1 and alpha 2-globin cDNAs were dot-blotted and further hybridized to synthetic oligonucleotides encompassing either the normal or the mutated sequences. The generated signals corresponded specifically to one of the two alpha-globin genes. Using this approach the alpha-globin structural mutants J-Buda and G-Pest were found to be encoded by the alpha 2 and the alpha 1-globin genes, respectively. Furthermore, the exact nucleotide changes were determined. We propose this technique to serve as a simple and definitive method for assigning alpha-globin structural mutants.
authors.affiliation: Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology, Heraklion, Greece.
authors.forename: A
authors.initials: A
authors.lastname: Mamalaki
authors.forename: M
authors.initials: M
authors.lastname: Horanyi
authors.forename: J
authors.initials: J
authors.lastname: Szelenyi
authors.forename: N K
authors.initials: NK
authors.lastname: Moschonas
mesh_terms: D001483:Base Sequence; D002874:Chromosome Mapping; D004247:DNA / Q000096:biosynthesis*; D004252:DNA Mutational Analysis; D005784:Gene Amplification; D005796:Genes / Q000235:genetics*; D005914:Globins / Q000235:genetics*; D006801:Humans; D015151:Immunoblotting; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000235:genetics
chemical_list: D015345:Oligonucleotide Probes; D005914:Globins; D004247:DNA