Production of Hageman factor (human FXII) in simian cells by transient expression of its cDNA.

Alessandro Aiuti; Monica Rinaldi; Claudia La Porta; Antonio Fantoni; Franca Citarella; M. Brunori

doi:10.1007/BF03001753

Production of Hageman factor (human FXII) in simian cells by transient expression of its cDNA.

Translated title of the contribution: Produzione di fattore di Hageman (FXII umano) in cellule di scimmia mediante espressione transiente del suo cDNA

Alessandro Aiuti, Monica Rinaldi, Claudia La Porta, Antonio Fantoni, Franca Citarella, M. Brunori

IRCCS Ospedale San Raffaele

Research output: Contribution to journal › Article › peer-review

Abstract

- Human coagulation factor XII (FXII) is a serine protease involved in a wide variety of functions, including blood coagulation, fibrinolysis, bradykinin production and complement activation. FXII consists of several autonomous structural regions similar to those present in other serine proteases. With the intent of assessing the contribution of each structural domain to the biological properties of the enzyme, we have built an experimental model that allows the expression of FXII cDNA in eukaryotic cells. The fulll-ength FXII cDNA was constructed and cloned into the shuttle vector pSVL under the SV40 promoter control. Simian cells (COS) were transfected with either the recombinant or the wild-type vector. At different times after transfection the cells and the conditioned medium were harvested and tested for the production and secretion of the recombinant FXII. FXII cDNA transcription and translation were demonstrated by Northern transfer, indirect immunofluorescence and Western blot, while the functional activity of the recombinant protein was proved by a coagulation assay.

Translated title of the contribution	Produzione di fattore di Hageman (FXII umano) in cellule di scimmia mediante espressione transiente del suo cDNA
Original language	Italian
Pages (from-to)	91-97
Number of pages	7
Journal	Rendiconti Lincei
Volume	1
Issue number	1
DOIs	https://doi.org/10.1007/BF03001753
Publication status	Published - Mar 1990

ASJC Scopus subject areas

Engineering(all)

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@article{13fef8a3c4c345779fee97da6fb62d43,

title = "Production of Hageman factor (human FXII) in simian cells by transient expression of its cDNA.",

abstract = "- Human coagulation factor XII (FXII) is a serine protease involved in a wide variety of functions, including blood coagulation, fibrinolysis, bradykinin production and complement activation. FXII consists of several autonomous structural regions similar to those present in other serine proteases. With the intent of assessing the contribution of each structural domain to the biological properties of the enzyme, we have built an experimental model that allows the expression of FXII cDNA in eukaryotic cells. The fulll-ength FXII cDNA was constructed and cloned into the shuttle vector pSVL under the SV40 promoter control. Simian cells (COS) were transfected with either the recombinant or the wild-type vector. At different times after transfection the cells and the conditioned medium were harvested and tested for the production and secretion of the recombinant FXII. FXII cDNA transcription and translation were demonstrated by Northern transfer, indirect immunofluorescence and Western blot, while the functional activity of the recombinant protein was proved by a coagulation assay.",

keywords = "COS cells, Factor XII, Transient expression",

author = "Alessandro Aiuti and Monica Rinaldi and Porta, {Claudia La} and Antonio Fantoni and Franca Citarella and M. Brunori",

year = "1990",

month = mar,

doi = "10.1007/BF03001753",

language = "Italian",

volume = "1",

pages = "91--97",

journal = "ATTI Della Accademia Nazionale Dei Lincei Rendiconti Lincei Scienze Fisiche E Naturali",

issn = "2037-4631",

publisher = "Accademia Nazionale dei Lincei",

number = "1",

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TY - JOUR

T1 - Production of Hageman factor (human FXII) in simian cells by transient expression of its cDNA.

AU - Aiuti, Alessandro

AU - Rinaldi, Monica

AU - Porta, Claudia La

AU - Fantoni, Antonio

AU - Citarella, Franca

AU - Brunori, M.

PY - 1990/3

Y1 - 1990/3

N2 - - Human coagulation factor XII (FXII) is a serine protease involved in a wide variety of functions, including blood coagulation, fibrinolysis, bradykinin production and complement activation. FXII consists of several autonomous structural regions similar to those present in other serine proteases. With the intent of assessing the contribution of each structural domain to the biological properties of the enzyme, we have built an experimental model that allows the expression of FXII cDNA in eukaryotic cells. The fulll-ength FXII cDNA was constructed and cloned into the shuttle vector pSVL under the SV40 promoter control. Simian cells (COS) were transfected with either the recombinant or the wild-type vector. At different times after transfection the cells and the conditioned medium were harvested and tested for the production and secretion of the recombinant FXII. FXII cDNA transcription and translation were demonstrated by Northern transfer, indirect immunofluorescence and Western blot, while the functional activity of the recombinant protein was proved by a coagulation assay.

AB - - Human coagulation factor XII (FXII) is a serine protease involved in a wide variety of functions, including blood coagulation, fibrinolysis, bradykinin production and complement activation. FXII consists of several autonomous structural regions similar to those present in other serine proteases. With the intent of assessing the contribution of each structural domain to the biological properties of the enzyme, we have built an experimental model that allows the expression of FXII cDNA in eukaryotic cells. The fulll-ength FXII cDNA was constructed and cloned into the shuttle vector pSVL under the SV40 promoter control. Simian cells (COS) were transfected with either the recombinant or the wild-type vector. At different times after transfection the cells and the conditioned medium were harvested and tested for the production and secretion of the recombinant FXII. FXII cDNA transcription and translation were demonstrated by Northern transfer, indirect immunofluorescence and Western blot, while the functional activity of the recombinant protein was proved by a coagulation assay.

KW - COS cells

KW - Factor XII

KW - Transient expression

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U2 - 10.1007/BF03001753

DO - 10.1007/BF03001753

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JO - ATTI Della Accademia Nazionale Dei Lincei Rendiconti Lincei Scienze Fisiche E Naturali

JF - ATTI Della Accademia Nazionale Dei Lincei Rendiconti Lincei Scienze Fisiche E Naturali

SN - 2037-4631

IS - 1

ER -

Production of Hageman factor (human FXII) in simian cells by transient expression of its cDNA.

Abstract

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