Production, purification and biological properties of an Escherichia coli-derived recombinant porcine alpha interferon.
Recombinant plasmids for intracellular synthesis of mature porcine interferon alpha 1 (IFN-alpha 1) in Escherichia coli were constructed. High amounts of antiviral activity were obtained [up to 4 x 10(5) international units (IU) per ml of bacterial culture]. Recombinant porcine IFN-alpha 1 (rIFN-alpha 1) was purified to homogeneity by monoclonal antibody immunoaffinity and was found to have the expected Mr (17.5K) and N-terminal sequence (except for the apparent lack of an N-terminal methionine). Its specific antiviral activity was 5 x 10(7) to 10 x 10(7) IU/mg MDBK cells. In vitro biological properties of this purified rIFN-alpha 1 were compared to those of virus-induced porcine leukocyte interferon: the two interferons shared similar antigenic determinants and had the same ability to induce a cytocidal effect on primary cultures of pig kidney epithelial cells. However, rIFN-alpha 1 was at least six times more active in inducing an antiviral state on homologous porcine cells. These properties are discussed in the light of a possible in vivo use of the purified recombinant molecule.
Institut National de la Recherche Agronomique, Centre de Recherches de Jouy-en-Josas, France.
F
F
Lefèvre
R
R
L'Haridon
F
F
Borras-Cuesta
C
C
La Bonnardière
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D002470:Cell Survival; D003588:Cytopathogenic Effect, Viral; D000939:Epitopes; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007370:Interferon Type I / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D008969:Molecular Sequence Data; D010957:Plasmids; D011994:Recombinant Proteins; D013552:Swine; D014721:Vesicular stomatitis Indiana virus / Q000502:physiology
D000939:Epitopes; D007370:Interferon Type I; D011994:Recombinant Proteins