title: Quantitative evaluation of mammalian skeletal muscle as a heterologous protein expression system.
abstract: The production of mammalian proteins in sufficient quantity and quality for structural and functional studies is a major challenge in biology. Intrinsic limitations of yeast and bacterial expression systems preclude their use for the synthesis of a significant number of mammalian proteins. This creates the necessity of well-identified expression systems based on mammalian cells. In this paper, we demonstrate that adult mammalian skeletal muscle, transfected in vivo by electroporation with DNA plasmids, is an excellent heterologous mammalian protein expression system. By using the fluorescent protein EGFP as a model, it is shown that muscle fibers express, during the course of a few days, large amounts of authentic replicas of transgenic proteins. Yields of approximately 1mg/g of tissue were obtained, comparable to those of other expression systems. The involvement of adult mammalian cells assures an optimal environment for proper protein folding and processing. All these advantages complement a methodology that is universally accessible to biomedical investigators and simple to implement.
authors.affiliation: Department of Physiology, David Geffen School of Medicine, UCLA, Los Angeles, CA, USA.
authors.forename: Marino
authors.initials: M
authors.lastname: DiFranco
authors.forename: Patricia
authors.initials: P
authors.lastname: Neco
authors.forename: Joana
authors.initials: J
authors.lastname: Capote
authors.forename: Pratap
authors.initials: P
authors.lastname: Meera
authors.forename: Julio L
authors.initials: JL
authors.lastname: Vergara
mesh_terms: D000818:Animals; D003001:Cloning, Molecular / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D006614:Hindlimb; D035002:Lower Extremity; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008958:Models, Molecular; D009124:Muscle Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D018482:Muscle, Skeletal / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D009124:Muscle Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins