CLONING AND CHARACTERIZATION OF THE BOVINE INTERLEUKIN-2-GENE

摘要

A chemically-synthesized oligonucleotide composing a portion ofthe nucleotide sequence of the human IL-2 is employed as a probe to isolatethegene coding for human IL-2. The human IL-2 gene is selected from a cDNAlibrary prepared from RNA produced by mitogen-stimulated Jurkat cells.Double-stranded eDNA is prepared from polyadenylated RNA extracted frombovine cells thought to produce interleukin-2. Such cDNA is inserted within aplasmid vector and the recombinant plasmid employed to transform hosts.Plasmid DNA, prepared from pools of the transformed hosts, is hybridized withaprobe composed of a large portion of the coding sequence of the human IL-2gene. Pools of host cells that provide a positive signal to the human eDNAprobeare identified, subdivided, and rescreened until a single positive colony isidentified. Bovine plasmid cDNA is prepared from this colony, and the bIL-2gene is sequenced. The plasmid DNA is employed to express recombinant bovineIL-2 in yeast and bacterial expression systems, with the expressed bovine IL-2purified to homogeneity by one or more reverse phase high-performance liquidchrom atography procedures.