Miyatake S, Yokota T, Lee F, Arai K
Proc Natl Acad Sci U S A. 1985 Jan;82(2):316-20. doi: 10.1073/pnas.82.2.316.
We have isolated the mouse interleukin 3 (IL-3) gene from a mouse sperm DNA library based on homology with the mouse mast-cell growth-factor (MCGF) cDNA sequence. The nucleotide sequence determined for the gene and its flanking regions reveals that the IL-3 gene is composed of four introns and five exons. The nucleotide sequence of the exons agrees with that determined for the MCGF cDNA. A "TATA"-like sequence preceded by a G + C-rich region is found in the 5' flanking region. At the 3' region of the second intron are nine repeats of a closely related 14-base-pair (bp) sequence. These repeated sequences share extensive homology with a 20-bp repeated sequence found in the human genome, which was shown to have enhancer activity. Eight out of nine repeats form a 73-bp duplicated sequence and each 73-bp repeat contains sequences homologous to the core sequence suggested for enhancer elements. These sequences may play a role in the expression of the IL-3 gene in concanavalin A- or antigen-stimulated T lymphocytes. Stable transformants of L cells generated by co-transformation of the IL-3 gene with pSV2neo constitutively express MCGF activity indicating that the isolated gene is functional in vivo.
我们基于与小鼠肥大细胞生长因子(MCGF)cDNA序列的同源性,从小鼠精子DNA文库中分离出了小鼠白细胞介素3(IL-3)基因。对该基因及其侧翼区域测定的核苷酸序列显示,IL-3基因由四个内含子和五个外显子组成。外显子的核苷酸序列与MCGF cDNA测定的序列一致。在5'侧翼区域发现了一个富含G + C区域的类似“TATA”的序列。在第二个内含子的3'区域有九个紧密相关的14碱基对(bp)序列的重复。这些重复序列与人类基因组中发现的一个20 bp重复序列具有广泛的同源性,该序列已被证明具有增强子活性。九个重复中的八个形成了一个73 bp的重复序列,每个73 bp重复序列都包含与增强子元件建议的核心序列同源的序列。这些序列可能在伴刀豆球蛋白A或抗原刺激的T淋巴细胞中IL-3基因的表达中起作用。通过将IL-3基因与pSV2neo共转化产生的L细胞稳定转化体组成性地表达MCGF活性,表明分离的基因在体内具有功能。