title: Expression of rat neurofilament proteins NF-L and NF-M in transfected non-neuronal cells.
abstract: Two cDNA clones fully encoding the rat neurofilament proteins NF-L and NF-M were subcloned into eukaryotic expression vectors behind the strong constitutive viral promoters from SV40 and Rous sarcoma viruses. Transient transfection of L tk- and Cos cell lines with these expression constructs resulted in cells expressing the neurofilament proteins in an intermediate filament-type pattern. Additionally, a putative juxtanuclear organizing center or region was observed in the transfected cells, most noticeable shortly after the transfection procedure. Stable transfections were performed on mouse L tk- and Swiss 3T6 cells using NF-L and NF-M constructs bearing an SV40 early promoter driven neomycin selectable marker. Although G418-resistant clones were recovered with both the NF-L and the NF-M constructs, only clones expressing immunofluorescently stainable amounts of NF-M were detected and established. Immunoelectron microscopic analysis revealed NF-M and vimentin proteins to be colocalized on the same intermediate filaments.
authors.affiliation: Department of Pharmacology, New York University Medical Center, NY.
authors.forename: S S
authors.initials: SS
authors.lastname: Chin
authors.forename: R K
authors.initials: RK
authors.lastname: Liem
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D003001:Cloning, Molecular; D003078:Colchicine / Q000494:pharmacology; D003599:Cytoskeleton / Q000032:analysis*; D004247:DNA / Q000235:genetics; D005455:Fluorescent Antibody Technique; D007381:Intermediate Filament Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D007382:Intermediate Filaments / Q000032:analysis* / Q000648:ultrastructure; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D016900:Neurofilament Proteins; D010957:Plasmids; D051381:Rats; D013997:Time Factors; D014162:Transfection; D014746:Vimentin / Q000032:analysis
chemical_list: D007381:Intermediate Filament Proteins; D016900:Neurofilament Proteins; D014746:Vimentin; C044882:neurofilament protein L; D004247:DNA; D003078:Colchicine