RESUMEN
Vigilin is an abundant, highly conserved, ubiquitous protein containing 15 related, but non-identical, K-homolous nucleic acid binding domains. The construction, transfection and production of recombinant vigilin in mammalian expression system were investigated. The whole length of vigilin was amplified by polymerase chain reaction [PCR] and ligated to pCEP-PU vector. The recombinant construct pCEP-PU with vigilin was produced and transfected into Human embryonic kidney cells in a specific culture medium. The secreted recombinant vigilin was purified from the serum-free medium to homogeneity by affinity chromatography. The conditioned and purified media were tested for the presence of vigilin by sodium dodecyl sulfate polyacrylamide gel electrophoresis [SDS-PAGE] and immunoblotting with specific antibody. An immunoreactive band with an apparent molecular mass of approximately 140 kDa was detected. Immunofluorescence staining of transfected cells with vigilin demonstrated that recombinant vigilin molecules are localized in the nucleus and cytoplasm. In summary, the purified recombinant vigilin will facilitate future studies that address the structure and function of vigilin