Stable Escherichia coli-Clostridium acetobutylicum shuttle vector for secretion of murine tumor necrosis factor alpha

J Theys; S Nuyts; W Landuyt; L Van Mellaert; C Dillen; M Bohringer; P Durre; P Lambin; J Anne

doi:10.1128/AEM.65.10.4295-4300.1999

Stable Escherichia coli-Clostridium acetobutylicum shuttle vector for secretion of murine tumor necrosis factor alpha

J Theys, S Nuyts, W Landuyt, L Van Mellaert, C Dillen, M Bohringer, P Durre, P Lambin, J Anne^*

^*Corresponding author for this work

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

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Abstract

Recombinant plasmids were constructed to secrete mouse tumor necrosis factor alpha (mTNF-alpha) from Clostridium acetobutylicum, The shuttle plasmids contained the clostridial endo-beta 1,4-glucanase (eglA) promoter and signal sequence that was fused in frame to the mTNF-alpha cDNA, The construction was first tested in Escherichia coli and then introduced in C. acetobutylicum DSM792 by electroporation. Controls confirmed the presence and stability of the recombinant plasmids in this organism. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and an in vitro cytotoxic assay were used to monitor expression and secretion of mTNF-a during growth. Significant levels of biologically active mTNF-alpha were measured in both lysates and supernatants. The present report deals with investigations on the elaboration of a gene transfer system for cancer treatment using anaerobic bacteria.

Original language	English
Pages (from-to)	4295-4300
Number of pages	6
Journal	Applied and Environmental Microbiology
Volume	65
Issue number	10
DOIs	https://doi.org/10.1128/AEM.65.10.4295-4300.1999
Publication status	Published - Oct 1999
Externally published	Yes

Keywords

STREPTOMYCES-LIVIDANS
ANAEROBIC-BACTERIA
DELIVERY SYSTEM
GENE-THERAPY
INVIVO
CELLS
ELECTROPORATION
COLONIZATION
EXPRESSION
BACILLUS

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10.1128/AEM.65.10.4295-4300.1999

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title = "Stable Escherichia coli-Clostridium acetobutylicum shuttle vector for secretion of murine tumor necrosis factor alpha",

abstract = "Recombinant plasmids were constructed to secrete mouse tumor necrosis factor alpha (mTNF-alpha) from Clostridium acetobutylicum, The shuttle plasmids contained the clostridial endo-beta 1,4-glucanase (eglA) promoter and signal sequence that was fused in frame to the mTNF-alpha cDNA, The construction was first tested in Escherichia coli and then introduced in C. acetobutylicum DSM792 by electroporation. Controls confirmed the presence and stability of the recombinant plasmids in this organism. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and an in vitro cytotoxic assay were used to monitor expression and secretion of mTNF-a during growth. Significant levels of biologically active mTNF-alpha were measured in both lysates and supernatants. The present report deals with investigations on the elaboration of a gene transfer system for cancer treatment using anaerobic bacteria.",

keywords = "STREPTOMYCES-LIVIDANS, ANAEROBIC-BACTERIA, DELIVERY SYSTEM, GENE-THERAPY, INVIVO, CELLS, ELECTROPORATION, COLONIZATION, EXPRESSION, BACILLUS",

author = "J Theys and S Nuyts and W Landuyt and {Van Mellaert}, L and C Dillen and M Bohringer and P Durre and P Lambin and J Anne",

year = "1999",

month = oct,

doi = "10.1128/AEM.65.10.4295-4300.1999",

language = "English",

volume = "65",

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journal = "Applied and Environmental Microbiology",

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publisher = "American Society for Microbiology",

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TY - JOUR

T1 - Stable Escherichia coli-Clostridium acetobutylicum shuttle vector for secretion of murine tumor necrosis factor alpha

AU - Theys, J

AU - Nuyts, S

AU - Landuyt, W

AU - Van Mellaert, L

AU - Dillen, C

AU - Bohringer, M

AU - Durre, P

AU - Lambin, P

AU - Anne, J

PY - 1999/10

Y1 - 1999/10

N2 - Recombinant plasmids were constructed to secrete mouse tumor necrosis factor alpha (mTNF-alpha) from Clostridium acetobutylicum, The shuttle plasmids contained the clostridial endo-beta 1,4-glucanase (eglA) promoter and signal sequence that was fused in frame to the mTNF-alpha cDNA, The construction was first tested in Escherichia coli and then introduced in C. acetobutylicum DSM792 by electroporation. Controls confirmed the presence and stability of the recombinant plasmids in this organism. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and an in vitro cytotoxic assay were used to monitor expression and secretion of mTNF-a during growth. Significant levels of biologically active mTNF-alpha were measured in both lysates and supernatants. The present report deals with investigations on the elaboration of a gene transfer system for cancer treatment using anaerobic bacteria.

AB - Recombinant plasmids were constructed to secrete mouse tumor necrosis factor alpha (mTNF-alpha) from Clostridium acetobutylicum, The shuttle plasmids contained the clostridial endo-beta 1,4-glucanase (eglA) promoter and signal sequence that was fused in frame to the mTNF-alpha cDNA, The construction was first tested in Escherichia coli and then introduced in C. acetobutylicum DSM792 by electroporation. Controls confirmed the presence and stability of the recombinant plasmids in this organism. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and an in vitro cytotoxic assay were used to monitor expression and secretion of mTNF-a during growth. Significant levels of biologically active mTNF-alpha were measured in both lysates and supernatants. The present report deals with investigations on the elaboration of a gene transfer system for cancer treatment using anaerobic bacteria.

KW - STREPTOMYCES-LIVIDANS

KW - ANAEROBIC-BACTERIA

KW - DELIVERY SYSTEM

KW - GENE-THERAPY

KW - INVIVO

KW - CELLS

KW - ELECTROPORATION

KW - COLONIZATION

KW - EXPRESSION

KW - BACILLUS

U2 - 10.1128/AEM.65.10.4295-4300.1999

DO - 10.1128/AEM.65.10.4295-4300.1999

M3 - Article

SN - 0099-2240

VL - 65

SP - 4295

EP - 4300

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

IS - 10

ER -

Stable Escherichia coli-Clostridium acetobutylicum shuttle vector for secretion of murine tumor necrosis factor alpha

Abstract

Keywords

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