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Locus assignment of human alpha-globin structural mutants by selective enzymatic amplification of alpha 1 and alpha
A Mamalaki1, M Horanyi, J Szelenyi
1Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology, Heraklion, Greece.
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.