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Cloning of eukaryotic genes in single-strand phage vectors: the human interferon genes
Authors:Donald W. Bowden  Jen-i Mao  Tina Gill  Kathy Hsiao  Jay S. Lillquist  Douglas Testa  Gerald F. Vovis
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
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.
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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