Isolation of a novel interferon gene and expression thereof.

摘要

A novel mature human alpha-interferon gene is isolated from a human genomic library, said gene is sequenced, inserted into a recombinant expression vector by recombinant DNA techniques and is thereafter used to transform bacteria which then express the gene as determined by the presence of interferon activity. Novel methods and compositions which allow for the rapid screening and isolation of mammalian genes including low copy genes from genomic libraries employing short probes are also disclosed. The method of the present invention comprises synthesizing at least one DNA probe about 17 nucleotides in length and comprising a DNA sequence homologous to at least one DNA sequence contained within said gene, labeling the DNA probe and thereafter hybridizing the labeled DNA probe to said genomic library employing an improved method for hybridizing labeled DNA probes to recombinant vectors contained within a host cell and affixed to nitrocellulose filters, the improvement comprising filtering the labeled DNA probes prior to its addition to a hybridization solution, adding the labeled DNA probes to the hybridization solution, employing a device to simultaneously rotate and incubate filters during prehybridization and hybridization, and adding 10 percent dextran sulfate to prehybridization and hybridization solutions and thereafter identifying said gene in a manner consistent with the genomic library and label chosen. Two synthetic 17-mer DNA probes with the base sequence of 5 min CAGCCAGGATGGAGTCC3 min and 5 min CCTCCCAGGCACAAGGG3 min and methods for the use thereof in identification and isolation of interferon genes are provided.