Interleukin-2 Receptor �� Gene Structure And Function

Interleukin 2 (IL-2), a key regulator of the immune response, is secreted by activated T cells upon antigenic stimulation. Binding of IL-2 to its high-affinity receptor (IL-2R ) induces proliferation as well as functional differentiation of T and B lymphocytes and natural killer activation. The high affinity IL-2 receptor is comprised of three component chains, IL-2Rα, IL-2Rβ, and IL-2Rγ. IL-2Rβ and IL-2Rγ are present constitutively in resting lymphocytes. In contrast, IL-2Rα is expressed only following activation and it is the unique subunit which strictly bind with IL-2. Therefore, the interleukin 2 receptor α-chain plays a critical role in IL-2 responsiveness, thus explaining why expression of this chain is highly regulated.Homologous recombination is a powerful tool which has only recently become available for correcting or mutating the desired chromosomal locus. To establish the model of gene targeting in somatic cells and study the function of IL-2Rα gene in lymphocytes, we perform gene targeting of IL-2Rα in Jurkat cells and investigate the transcription regulation activity of the 1st intron of IL-2Rα gene.I. Establishment of cell cloning in semi-solid medium in suspension cell and the construction of Jurkat Tet-on Cell line.We successfully constructed Jurkat Tet-on cell line-rtTA(3) according to the protocol of hybridoma cloning culture using semi-solid medium of methyl cellulose. rtTA gene integrated into the chromosome of rtTA(3) cell line . These positive rtTA(3) cell line identified by Southern blot could express 30 folds luciferase highly while induced by 1 μg/ml of Dox.