title: Human ubiquitin C promoter based expression of erythropoietin in CHO K1 cell lines: a simple transfectants screening approach.
abstract: Erythropoietin (EPO), a glycoprotein hormone that regulates the production of erythrocytes in the human body, is of clinical importance in the treatment of anemia. Low expression levels of this recombinant hormone and time-consuming screening methods have made its commercial production expensive. Cloning of human EPO gene in a shuttle vector pUB6/V5-HisB driven by human ubiquitin C promoter and its transfection in CHO K1 cell lines by electroporation resulted in a moderate level of EPO expression. The limiting-dilution screening method required several months to obtain high expression stable transfectants but needed only short duration for selection in contrast to the present screening strategy. The supernatants of stably transfected cells were found to be biologically active by in vitro erythroid cluster forming activity.
authors.affiliation: Center for Biotechnology, Anna University, Guindy, Chennai 600025, India.
authors.forename: Murugan
authors.initials: M
authors.lastname: Surendarbabu
authors.forename: Sankaranarayanan
authors.initials: S
authors.lastname: Meenakshisundaram
mesh_terms: D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004351:Drug Resistance; D041905:Erythroid Cells / Q000166:cytology / Q000187:drug effects; D004921:Erythropoietin / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D051379:Mice; D009705:Nucleosides; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D014162:Transfection / Q000379:methods*; D025822:Ubiquitin C / Q000235:genetics*
chemical_list: D009705:Nucleosides; D011993:Recombinant Fusion Proteins; D025822:Ubiquitin C; D004921:Erythropoietin; C004500:blasticidin S