Production and characterization of recombinant equine prorelaxin.

摘要: Relaxin is a peptide hormone produced by wide variety of mammals. In the horse, placenta major source relaxin. Since pure equine relaxin difficult to obtain study its role in pregnant mare, objectives this were produce recombinant prorelaxin and characterize immunological biological activity. First, an gene cassette was transfected into immortalized bovine mammary epithelial (MAC-T) cells. Second, activity media conditioned MAC-T cells tested Western blotting quantified using homologous radioimmunoassay. Finally, bioactivity human monocyte cell line, THP-1, which exhibits rapid dose-dependent increase accumulation cAMP upon binding The results showed that media, concentrated 5x, yielded 4.11 +/- 0.81 ng/ml prorelaxin. addition, 19 kDa immunoreactive band, corresponding expected size prorelaxin, visualized SDS-PAGE. THP-1 incubated with (5x) from cells, presence forskolin (1 microM) isobutylmethylxanthine (50 microM), production over mock-transfected alone. conclusion, secreted both immunologically biologically active. This demonstrates first attempt important for further mare.