Generation of full-length cDNA recombinant vectors for the transient expression of human fibronectin in mammalian cell lines

In order to study the roles of the different alternatively spliced variants of human fibronectin (FN) as well as of its binding sites and structural domains in the processes of extracellular matrix assembly, cell adhesion, and cell migration, we constructed expression vectors coding for different hu...

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Detalles Bibliográficos
Publicado:	1991
Materias:	complementary dna fibronectin animal cell article cell adhesion expression vector extracellular matrix human mouse priority journal recombinant dna technology Animal Cloning, Molecular DNA Fibronectins Fluorescent Antibody Technique Gene Expression Genetic Vectors Human Mice Microinjections Support, Non-U.S. Gov't Transfection Mammalia
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00144827_v193_n2_p331_Dufour http://hdl.handle.net/20.500.12110/paper_00144827_v193_n2_p331_Dufour
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_00144827_v193_n2_p331_Dufour
record_format	dspace
spelling	paper:paper_00144827_v193_n2_p331_Dufour2023-06-08T14:37:16Z Generation of full-length cDNA recombinant vectors for the transient expression of human fibronectin in mammalian cell lines complementary dna fibronectin animal cell article cell adhesion expression vector extracellular matrix human mouse priority journal recombinant dna technology Animal Cloning, Molecular DNA Fibronectins Fluorescent Antibody Technique Gene Expression Genetic Vectors Human Mice Microinjections Support, Non-U.S. Gov't Transfection Mammalia In order to study the roles of the different alternatively spliced variants of human fibronectin (FN) as well as of its binding sites and structural domains in the processes of extracellular matrix assembly, cell adhesion, and cell migration, we constructed expression vectors coding for different human full-length FN polypeptides and deleted versions of these constructs. We expressed them transiently in mammalian cells by calcium phosphate transfection and microinjection techniques. While the deleted recombinants were expressed by both transfection and microinjection, the full-length recombinants could be only expressed by microinjection. Calcium phosphate transfection leads to the accumulation of recombinant FN in cytoplasmic vesicles of both matrix-forming and nonforming cells. On the contrary, microinjection leads to the accumulation of recombinant FN in cytoplasmic vesicles in cells that do not form a matrix, but to the rapid incorporation into the extracellular matrix of matrix-forming cells in addition to a cytoplasmic localization. Identical results were obtained when the FN signal and propeptides were replaced by those of E-cadherin. © 1991. 1991 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00144827_v193_n2_p331_Dufour http://hdl.handle.net/20.500.12110/paper_00144827_v193_n2_p331_Dufour
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	complementary dna fibronectin animal cell article cell adhesion expression vector extracellular matrix human mouse priority journal recombinant dna technology Animal Cloning, Molecular DNA Fibronectins Fluorescent Antibody Technique Gene Expression Genetic Vectors Human Mice Microinjections Support, Non-U.S. Gov't Transfection Mammalia
spellingShingle	complementary dna fibronectin animal cell article cell adhesion expression vector extracellular matrix human mouse priority journal recombinant dna technology Animal Cloning, Molecular DNA Fibronectins Fluorescent Antibody Technique Gene Expression Genetic Vectors Human Mice Microinjections Support, Non-U.S. Gov't Transfection Mammalia Generation of full-length cDNA recombinant vectors for the transient expression of human fibronectin in mammalian cell lines
topic_facet	complementary dna fibronectin animal cell article cell adhesion expression vector extracellular matrix human mouse priority journal recombinant dna technology Animal Cloning, Molecular DNA Fibronectins Fluorescent Antibody Technique Gene Expression Genetic Vectors Human Mice Microinjections Support, Non-U.S. Gov't Transfection Mammalia
description	In order to study the roles of the different alternatively spliced variants of human fibronectin (FN) as well as of its binding sites and structural domains in the processes of extracellular matrix assembly, cell adhesion, and cell migration, we constructed expression vectors coding for different human full-length FN polypeptides and deleted versions of these constructs. We expressed them transiently in mammalian cells by calcium phosphate transfection and microinjection techniques. While the deleted recombinants were expressed by both transfection and microinjection, the full-length recombinants could be only expressed by microinjection. Calcium phosphate transfection leads to the accumulation of recombinant FN in cytoplasmic vesicles of both matrix-forming and nonforming cells. On the contrary, microinjection leads to the accumulation of recombinant FN in cytoplasmic vesicles in cells that do not form a matrix, but to the rapid incorporation into the extracellular matrix of matrix-forming cells in addition to a cytoplasmic localization. Identical results were obtained when the FN signal and propeptides were replaced by those of E-cadherin. © 1991.
title	Generation of full-length cDNA recombinant vectors for the transient expression of human fibronectin in mammalian cell lines
title_short	Generation of full-length cDNA recombinant vectors for the transient expression of human fibronectin in mammalian cell lines
title_full	Generation of full-length cDNA recombinant vectors for the transient expression of human fibronectin in mammalian cell lines
title_fullStr	Generation of full-length cDNA recombinant vectors for the transient expression of human fibronectin in mammalian cell lines
title_full_unstemmed	Generation of full-length cDNA recombinant vectors for the transient expression of human fibronectin in mammalian cell lines
title_sort	generation of full-length cdna recombinant vectors for the transient expression of human fibronectin in mammalian cell lines
publishDate	1991
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00144827_v193_n2_p331_Dufour http://hdl.handle.net/20.500.12110/paper_00144827_v193_n2_p331_Dufour
_version_	1768542869210005504

Generation of full-length cDNA recombinant vectors for the transient expression of human fibronectin in mammalian cell lines

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