Man Il-2 Gene Cloning And Mammalian Cell Transfection Study

T lymphocytes are the critical cells involved in the regulation of immune responses and mediation of cytolytic activity. In order to dissect immune mechanisms, it is important to develop the technology for amplifying and even cloning the T cell subpo-pulations in vitro. Interleukin 2 ( IL-2 ), one of T cell growth factors, possesses multiple biological activities. The basic aim of the present study is to develop the technical system of effective transfer of human IL—2 cDNA into mammalian lymphocytes with stable expression of the gene product. This will initiate the studies of cloning T cell subpopuLations in vitro by gene transfe and deepening the research on regulatory mechanisms of gene expre ssion involved in T cell growth.Two DNA fragments were selectively amplified from the total RNA extracted from the PHA-stimulated human tonsilar cells by using the technique of polymerase chain reaction initiated by two sets of primers synthesized by DNA sythesizer. These DNA fragments were identified by using Southern blot and DNA sequence analysis. A long fragment, about 5Q0bp in lenghth, is the cDNA encoding for the whole IL-2 amino acid sequence including the signal peptide, and another one, about 400 bp, encodes for mature IL-2. The sequence obtained was idetical to those reported in the literatur The shorter IL-2 cDNA was inserted in proper orientation at the...