Open AccessSite-specific mutagenesis of the human fibroblast interferon gene.
Author(s) -
D F Mark,
Shi-Da Lu,
A A Creasey,
R Yamamoto,
L S Lin
Publication year - 1984
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.81.18.5662
Subject(s) - interferon , microbiology and biotechnology , mutagenesis , site directed mutagenesis , cysteine , biology , primer (cosmetics) , fibroblast , oligonucleotide , gene , amino acid , biochemistry , chemistry , virology , mutation , in vitro , mutant , organic chemistry , enzyme
Human fibroblast interferon has three cysteine residues, located at amino acid positions 17, 31, and 141. Using the technique of site-specific mutagenesis with a synthetic oligonucleotide primer, we changed the codon for cysteine-17 to a codon for serine. The resulting interferon, IFN-beta Ser-17, retains the antiviral, natural killer cell activation, and antiproliferative activities of native fibroblast interferon. The purified IFN-beta Ser-17 protein has an antiviral specific activity of 2 X 10(8) units/mg, similar to that of purified native fibroblast interferon. In addition, the purified protein is stable to long-term storage at -70 degrees C.