The bovine IFN-omega 1 is biologically active and secreted at high levels in the yeast Pichia pastoris.
The gene coding for bIFN-omega 1 was isolated from bovine genomic DNA by polymerase chain reaction (PCR). Recombinant bIFN-omega 1 was expressed in the yeast Pichia pastoris and high levels of the recombinant protein (0.4 mg ml-1) were secreted to the culture media. The obtained bIFN-omega 1 showed a cross-species antiviral activity on four mammalian cell lines of calf, pig, hamster and human origin, but this activity was absent on Madin-Darby canine kidney (MDCK) cells. A delivery carrier was developed to permit a better release of bIFN-omega 1. When compared with a control group, an increase in 6 days in the corpus luteum lifespan was obtained in cyclic ewes following three interferon (IFN) intrauterine administrations on days 9, 10 and 11 post-estrus. In summary, these results demonstrated for the first time that biologically active recombinant bIFN-omega 1 was highly secreted by P. pastoris showing antiviral activity in different cell lines and an antiluteolytic effect in cyclic ewes, with no detrimental effects on the animals.
Rodríguez
M
M
Division of Mammalian Cell Genetics, Centro de Ingeniería Genética y Biotecnología, Havana, Cuba.
Martínez
V
V
Alazo
K
K
Suárez
M
M
Redondo
M
M
Montero
C
C
Besada
V
V
de la Fuente
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D002417:Cattle; D002460:Cell Line; D003338:Corpus Luteum / Q000502:physiology; D006224:Cricetinae; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D015870:Gene Expression; D006801:Humans; D007370:Interferon Type I / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011374:Progesterone / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D012756:Sheep; D013552:Swine; D014599:Uterus / Q000378:metabolism
D000998:Antiviral Agents; D007370:Interferon Type I; D011994:Recombinant Proteins; C063743:interferon omega 1; D011374:Progesterone