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Detection of complex alleles by direct analysis of DNA heteroduplexes |
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| Authors: | Rosa Sorrentino Carlo Iannicola Sandro Costanzi Alberto Chersi Roberto Tosi |
| Institution: | (1) Dipartimento di Biologia Cellulare e dello Sviluppo, Università  La Sapienza , Roma, Italy;(2) Instituto di Biologia Cellulare, CNR, Roma, Italy;(3) Cattedra di Genetica Medica, Dipartimento di Medicina Sperimentale, Università La Sapienza, Roma, Italy;(4) Istituto Regina Elena, Ripartizione di Oncologia Sperimentale, Roma, Italy;(5) Istituto di Biologia Cellulare, Viale Marx 43, 00137 Roma, Italy |
| Abstract: | DNA molecules derived from three alleles of the HLA-DRB3 locus and differing from each other at several nucleotide sites were denatured and cross-hybridized. Each allelic combination was found to generate a pair of heteroduplexes of different mobility. Their retardation as compared to homoduplexes was proportional to the number of mismatches. In each heteroduplexes pair the component possessing the highest number of Pyr-Pyr oppositions was the most retarded. The results are those predicted by a theoretical model implying a correlation between base-pair opening and bending of the DNA double helix. These observations introduce a new HLA typing method at the genomic level and indicate an experimental approach to the analysis of the superhelical DNA conformation as related to different types of base oppositions. |
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