Cloning and structure of the human interleukin 2 chromosomal gene.

Abstract

Southern hybridization using 32P‐labelled human interleukin 2 (IL2) cDNA probes revealed the existence of a single human IL2 gene. Five clones containing the human IL2 chromosomal gene were isolated from two different human DNA libraries cloned in either lambda Charon 4A or L47 phages. Analysis of the clones showed that they contained different, overlapping portions of human DNA which were derived from the same chromosomal segment. Restriction fragments which hybridized with labelled IL2 cDNA probes were subcloned into plasmid pUR250 and the sequence and organization of the IL2 gene was determined. It contains three introns, 90 bp, +/‐ 2400 bp and +/‐ 1900 bp in length, respectively. The organization of the genomic clone resembles that of another lymphokine, interferon‐gamma, but no clear homology was found by comparing either the coding sequence or the 5′‐ and 3′‐flanking sequences of the two genes.

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

“…The 6 bp (AAATTA) direct repeats are generated at this second process. As the nucleotide sequences of the 6 bp direct repeats are apparently more conserved than that of the 10 bp repeats, our observation supports the idea that the transposition of the Alu family took place earlier than that (3,8,21). The 'TATA' box is enclosed.…”

“…In ongoing investigations of the mechanisms of IL-2 induction, we cloned a human genomic DNA fragment containing the entire IL-2 gene (21). This gene is approximately 5 kb long and is split into four exons (3,8,12,21). Nucleotide sequence analysis of a part of the 5' flanking region ofthe IL-2 gene revealed a homology with that of the corresponding region of the human immune interferon gene (3,8,21).…”

Small-Molecule Inhibitors of IL-2/IL-2R: Lessons Learned and Applied

“…The gene for IL-2 was cloned in 1983 (Degrave et al 1983;Taniguchi et al 1983), and the crystal structure ( Fig. 3) determined in 1987 (Brandhuber et al 1987).…”

Expression of amplified cDNA sequences encoding human interleukin 2 in human cells.

“…7 and C1.14 were inserted singly between the Hindlll and Bglll sites of pSV2dhfr to yield pSV2-IL2 C1.7 and C1.14, respectively.…”

“…Since pSV2-IL2 C1.7 was more active than pSV2-IL2 C1.14 in producing IL-2 by the transient expression assay in COS-1 cells (Table I), a small fragment of pSV2-IL2 C1. 7 cleaved with Pvull and BamHlwas used to construct the plasmid PSDI (Fig. 1), the hybrid Table I HeLa-PSDI-R400 2, 700 containing both IL-2 and DHFR cDNAs, which expressed IL-2 in COS-1 cells (Table I).…”