Renaturation rate studies of a single family of interspersed repeated sequences in human deoxyribonucleic acid

Abstract

We have investigated the renaturation kinetics of a single family of cloned interspersed repeated sequences isolated from human DNA. Cross-renaturation studies of individual cloned sequences reveal heterogeneity in both the renaturation rate and the thermal stability of heteroduplexes formed from members of this family of sequences. However, cloned members of this family all renature with approximately the same number of copies in the human genome, demonstrating that they are a single family of sequences by the criterion of DNA renaturation kinetics. When a single cloned member of the family is renatured with total human DNA as a function of temperature, the thermal stability of the renatured heteroduplexes is found to be independent of the renaturation temperature over the range 25 degrees C below to 4 degrees C above the melting temperature. Further, the number of genomic copies with which this cloned family member reacts is also independent of the renaturation temperature from 25 degrees C below up to the melting temperatures. These observations demonstrate a remarkable degree of homogeneity in the evolutionary sequence divergence of members of this family. These results also demonstrate that renaturation kinetics can accurately measure the number of genomic copies of interspersed repeated DNA sequences.

HLA‐DRB9— Possible Remnant of an Ancient Functional DRB Subregion

“…not described previously at homologous positions). 'Allelism' of the elements is indicated by sharing of diagnostic substitutions [21,22] and sharing of flanking inverted repeats which presumably arose during the insertion of the Alu element at a given chromosomal site [23]. Thus, of the three Alu elements, one is an allele of Alu 50 which is present in intron 1 of all human DRB genes [24].…”

Alu elements in human growth hormone receptor gene 5' untranslated region exons

“…Alu sequences, the largest family of short interspersed repetitive elements (SINEs) in primates, can be found inserted on average every 4 kb of genomic DNA, accounting for 10% of the human genome (Rinehart et al 1981, IHGSC 2001. A typical Alu element is 300 nt, composed of two homologous but distinct subunits, right and left, derived originally from the 7SL RNA gene (Labuda et al 1995).…”

Single DNA marker generated by “YAC-Alu PCR” that is end-specific

“…Thus, the frequency of the generation of the products of PCR from YACs can be expected to be lower in the case of YACs that contain Alu-poor domains. To circumvent these difficulties, we recommend the use of primers that correspond to other repeated sequences, such as the L1 repeated sequences and GT: AC families (Hwu et aL, 1986;Rinehart et aL, 1981). The inverse PCR technology (Ochman et al, 1988;Triglia et al, 1988;Silverman et aL, 1989) and the linker-specific amplification technique (Riley et al, 1990) are other useful tools for preparation of linking probes.…”