<b>ORGANIZATION OF THE DNA REGIONS FLANKING THE HUMAN INTERLEUKIN 2 </b><b>GENE </b>

Abstract

The organization of an 18 kilobase (kb) nuclear DNA fragment containing the entire human interleukin 2 (IL-2) gene was investigated by Southern blot analysis and DNA sequencing. A 6 kb cluster of the Kpnl families is located 3 kb upstream from the 5' end ofthe IL-2 gene. The members ofthe cluster and their arrangements are identical to previous determinations of the Kpnl cluster present in the human ,6'-globin gene region. One of the Alu family and a d(A-C)14d(T-C)24 sequence are linked and are located 2.5 kb downstream from the 3' end ofthe IL-2 gene. This Alu sequence is flanked with l0 basepair (bp) direct repeats, and the Alu and the homocopolymer sequences seem to form a composite movable element, as deduced from the finding that this unit is also flanked with 6 bp direct repeats. Tissue-specific DNA rearrangements were examined in the 5' flanking region of the human IL-2 gene.

Mapping of the IDDM Locus Idd3 to a 0.35-cM Interval Containing the Interleukin-2 Gene

“…Because the serine at position 6 is conserved between the two species, we sequenced exon 1 of the IL2 gene in 103 individuals to determine if the same polymorphism exists in humans. Although polymorphisms have previously been reported (20,21) in exon 1 of the IL2 gene, the only polymorphism we observed in 206 Caucasian and black chromosomes was a T to G substitution at nucleotide position 742. Of the 103 individuals sequenced, 21 were homozygous for the T allele, 57 were homozygous for the G allele, and 25 were heterozygous.…”

“…Briefly, YAC vectorette libraries were amplified by interspersed repetitive element (IRE)/bubble PCR using the conditions of Munroe et al (13). PCR products were annealed with biotinylated (TG) 20 and (AG) 20 oligonucleotides to enrich for products containing microsatellites. Following capture on streptavidin-coated magnetic beads (Dynal, Oslo, Norway), the IRE/bubble products were reamplified, cloned into pAMPIO (Life Technologies), and transformed into DH5a competent cells (Life Technologies).…”