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Nucleic Acids Research, 1984, Vol. 12, No. 2 1227-1242
© 1984

MOLECULAR BIOLOGY

Structure and expression in Escherichia coil of a cloned rat interferon-{alpha} gene

R. Dijkema, P. Pouwels, A. de Reus* and H. Schellekens*

Medical Biological Laboratory TNO P.O. Box 45, 2280 AA *primate Centre TNO P.O. Box 5815, 2280 HV Rijswijk, The Netherlands

Received October 21, 1983. Accepted November 28, 1983.

DNA synthesized by in vitro transcription on rat interferon (IFN) mRNA has been cloned and amplified as recombinant DNA. The nucleotide sequence of these rat IFN cDNA clones revealed i. the partial presence of the coding region of the gene and ii all cDNA clones were derived from the same subtype of rat IFN-{alpha} mRNA. Purified inserted fragments were used as a hybridisation probe against chromosomal "Southern blots" to show that at least twelve rat IFN-{alpha}-related sequences are present in the genome. A {lambda}-linked rat gene library was screened with the cDNA probes, resulting in an equivalent number of rat IFN-{alpha}-related hybrid phages. By use of a 3'-noncoding region as a probe, the chrornosomal counterpart of the cDNA clones could be detected and the nucleotide sequence of its coding region has been determined. Expression of the coding region in E. coli yielded biologically active IFN, when tested for in vitro or in vivo antiviral activity.


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