| 摘要 |
A chemically-synthesized oligonucleotide composing a portion of the nucleotide sequence of human IFN- gamma is employed as a probe to isolate the gene coding for human lFN- gamma from a cDNA library prepared form RNA derived from peripheral blood leukocytes. Double stranded cDNA is prepared from the polyadenylated RNA extracted from bovine cells thought to produce IFN- gamma . Such cDNA is inserted within a plasmid vector and the recombinant plasmid employed to transform hosts. Plasmid DNA, prepared from the pools of the transformed hosts, is hybridized with the human IFN- gamma gene probe. Pools of host cells that provide a positive signal to the human cDNA probe are identified, subdivided and rescreened until a single positive colony is identified. Bovine plasmid cDNA is prepared from this colony, and the bIFN- gamma gene is sequenced. The coding region for the bIFN- gamma gene is inserted into an expression vector for production of functional bIFN- gamma . |
