Cloning of eukaryotic genes in single-strand phage vectors: the human interferon genes |
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Authors: | Donald W. Bowden Jen-i Mao Tina Gill Kathy Hsiao Jay S. Lillquist Douglas Testa Gerald F. Vovis |
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Affiliation: | 1. Collaborative Research, Inc., 128 Spring St., Lexington, MA 02173 U.S.A. Tel. (617) 861-9700;1. Interferon Sciences, Inc., 783 Jersey Avenue, New Brunswick, NJ 08901 U.S.A. Tel. (201) 249-3232 |
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Abstract: | Using oligonucleotide probes with defined sequences, we have selected clones from a human lymphocyte cDNA library which represent human leukocyte (HuIFN-α) and fibroblast (HuIFN-β) interferon gene sequences. Double-stranded f1 phage DNA was used as the vector for initial cloning of cDNA. Clones carrying interferon gene sequences were identified by hybridization with the oligonucleotide probes. The same oligonucleotide probes were used as primers for dideoxy chain termination sequencing of the clones. One HuIFN-α clone, 201, has a nucleotide sequence different from published HuIFN-α sequences. Under control of the lacUV5 promoter, the 201 gene has been used to express biologically active HuIFN-α in Escherichia coli. |
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Keywords: | Recombinant DNA bacteriophage f1 leukocyte fibroblast cDNA DNA sequencing AMV, avian myeloblastosis virus bp, base pairs cDNA, DNA complementary to mRNA CPE, cytopathic effect DTT, dithiothreitol HuIFN, human interferon (α, leukocyte β, fibroblast) RF, replicative form of phage f1 DNA SDS, sodium dodecyl sulfate TY broth, see MATERIALS AND METHODS, section a [ ], indicates plasmid carrier state |
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