Length and sequence variation in D7S22 (g3) alleles studied by high resolution length measurements and nucleotide sequencing

In a study of DNA sequence and length variation in the repeat array of small D7S22 alleles, 100 alleles typed as the common 14 repeat allele (14R) and 92 rare ones were selected for further characterization. A polymerase chain reaction (PCR) based allele length measurement method revealed a discontinuous distribution of alleles. The 92 rare alleles were grouped by their number of repeats. All, except four 6R alleles were distributed within the 11R-19R allele groups. The 14Rs revealed no further length variation while 7 out of the 92 rare alleles showed small length deviations from the other alleles within their respective groups. Nucleotide sequencing of the repeat array was performed in 17 alleles selected from each of the nine allele groups. The micro length variation within allele groups was caused by the presence of either 33, 36 or 37 bp repeats in given positions. A comparison of three 14Rs revealed no further sequence variation between these. Nine out of the fourteen repeats in the 14R differed in sequence and/or size. Based on this difference the repeat array sequence was converted into a code of different variant repeats. The 6R showed a variant repeat code quite unlike that of the 14R, while the encoded allele structure of the other rare alleles suggested that most of them may have evolved from a 14R allele by deletion or duplication of repeat units. Nucleotide sequencing of progenitor and mutant in a D7S22 de novo mutation as well as typing in a polymorphic site near the repeat array suggested that the event was an intra-allelic deletion of exactly three repeats. The present findings indicate that the 14R is ancestral to most rare small alleles, and that mutations in small alleles most often are intra-allelic events leading to a change in bp size equal to an integer number of repeats.