Biological and immunological characterization of a cloned cholera toxin-like enterotoxin from Salmonella typhimurium

Rajendra Prasad; Ashok K. Chopra; Johnny W. Peterson; Roser Pericas; Clifford W. Houston

doi:10.1016/0882-4010(90)90066-Y

Biological and immunological characterization of a cloned cholera toxin-like enterotoxin from Salmonella typhimurium

Rajendra Prasad, Ashok K. Chopra, Johnny W. Peterson, Roser Pericas, Clifford W. Houston

Microbiology And Immunology

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

27 Scopus citations

Abstract

A chromosomal DNA fragment, encoding an enterotoxin gene of Salmonella typhimurium Q1, was cloned into bacteriophage EMBL3 and plasmid vector pBR322. The recombinant clones λB8 and pC1 were identified using a synthetic oligonucleotide probe made to the B subunit region of the cholera toxin gene (ctx). Cell lysates of Escherichia coli VCS257[λB8] induced fluid secretion in rabbit intestinal loops, while lysates of E. coli DH5α[pC1] failed to elicit an enterotoxic response in this model. Both lysates and partially purified preparations elongated Chinese hamster ovary (CHO) cells, elevated cellular cAMP and PGE₂, and bound to ganglioside G_M1. The biological activity associated with the cloned enterotoxin was neutralized by monospecific antiserum to cholera toxin (CT). Immunoblots of pC1 and λB8 lysates probed with anti-CT, exhibited a 30 kDa protein similar to that of pJM17, which carried the ctx gene. Under non-dissociating conditions, anti-CT immunoblots of the same lysates revealed two proteins, one corresponding in size to the holotoxin and the other to CT-A. When analysed by DNA-directed protein synthesis in vitro, both pC1 and λB8 DNA expressed two unique proteins (30 and 11 kDa) similar to that of pJM17.

Original language	English (US)
Pages (from-to)	315-329
Number of pages	15
Journal	Microbial Pathogenesis
Volume	9
Issue number	5
DOIs	https://doi.org/10.1016/0882-4010(90)90066-Y
State	Published - Nov 1990

Keywords

ELISA
G ganglioside
PGE
Salmonella enterotoxin
cAMP
cholera toxin

ASJC Scopus subject areas

Microbiology
Infectious Diseases

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10.1016/0882-4010(90)90066-Y

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@article{4b9689b354134594bac64dc020fb09fa,

title = "Biological and immunological characterization of a cloned cholera toxin-like enterotoxin from Salmonella typhimurium",

abstract = "A chromosomal DNA fragment, encoding an enterotoxin gene of Salmonella typhimurium Q1, was cloned into bacteriophage EMBL3 and plasmid vector pBR322. The recombinant clones λB8 and pC1 were identified using a synthetic oligonucleotide probe made to the B subunit region of the cholera toxin gene (ctx). Cell lysates of Escherichia coli VCS257[λB8] induced fluid secretion in rabbit intestinal loops, while lysates of E. coli DH5α[pC1] failed to elicit an enterotoxic response in this model. Both lysates and partially purified preparations elongated Chinese hamster ovary (CHO) cells, elevated cellular cAMP and PGE2, and bound to ganglioside GM1. The biological activity associated with the cloned enterotoxin was neutralized by monospecific antiserum to cholera toxin (CT). Immunoblots of pC1 and λB8 lysates probed with anti-CT, exhibited a 30 kDa protein similar to that of pJM17, which carried the ctx gene. Under non-dissociating conditions, anti-CT immunoblots of the same lysates revealed two proteins, one corresponding in size to the holotoxin and the other to CT-A. When analysed by DNA-directed protein synthesis in vitro, both pC1 and λB8 DNA expressed two unique proteins (30 and 11 kDa) similar to that of pJM17.",

keywords = "ELISA, G ganglioside, PGE, Salmonella enterotoxin, cAMP, cholera toxin",

author = "Rajendra Prasad and Chopra, {Ashok K.} and Peterson, {Johnny W.} and Roser Pericas and Houston, {Clifford W.}",

note = "Funding Information: This work was supported by research grant R01 All 8401 from the National Institutes of Health . The authors thank M . Susman for editorial assistance during manuscript preparation and C . Jackson for performing the cAMP and PGE 2 assays .",

year = "1990",

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doi = "10.1016/0882-4010(90)90066-Y",

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T1 - Biological and immunological characterization of a cloned cholera toxin-like enterotoxin from Salmonella typhimurium

AU - Prasad, Rajendra

AU - Chopra, Ashok K.

AU - Peterson, Johnny W.

AU - Pericas, Roser

AU - Houston, Clifford W.

N1 - Funding Information: This work was supported by research grant R01 All 8401 from the National Institutes of Health . The authors thank M . Susman for editorial assistance during manuscript preparation and C . Jackson for performing the cAMP and PGE 2 assays .

PY - 1990/11

Y1 - 1990/11

N2 - A chromosomal DNA fragment, encoding an enterotoxin gene of Salmonella typhimurium Q1, was cloned into bacteriophage EMBL3 and plasmid vector pBR322. The recombinant clones λB8 and pC1 were identified using a synthetic oligonucleotide probe made to the B subunit region of the cholera toxin gene (ctx). Cell lysates of Escherichia coli VCS257[λB8] induced fluid secretion in rabbit intestinal loops, while lysates of E. coli DH5α[pC1] failed to elicit an enterotoxic response in this model. Both lysates and partially purified preparations elongated Chinese hamster ovary (CHO) cells, elevated cellular cAMP and PGE2, and bound to ganglioside GM1. The biological activity associated with the cloned enterotoxin was neutralized by monospecific antiserum to cholera toxin (CT). Immunoblots of pC1 and λB8 lysates probed with anti-CT, exhibited a 30 kDa protein similar to that of pJM17, which carried the ctx gene. Under non-dissociating conditions, anti-CT immunoblots of the same lysates revealed two proteins, one corresponding in size to the holotoxin and the other to CT-A. When analysed by DNA-directed protein synthesis in vitro, both pC1 and λB8 DNA expressed two unique proteins (30 and 11 kDa) similar to that of pJM17.

AB - A chromosomal DNA fragment, encoding an enterotoxin gene of Salmonella typhimurium Q1, was cloned into bacteriophage EMBL3 and plasmid vector pBR322. The recombinant clones λB8 and pC1 were identified using a synthetic oligonucleotide probe made to the B subunit region of the cholera toxin gene (ctx). Cell lysates of Escherichia coli VCS257[λB8] induced fluid secretion in rabbit intestinal loops, while lysates of E. coli DH5α[pC1] failed to elicit an enterotoxic response in this model. Both lysates and partially purified preparations elongated Chinese hamster ovary (CHO) cells, elevated cellular cAMP and PGE2, and bound to ganglioside GM1. The biological activity associated with the cloned enterotoxin was neutralized by monospecific antiserum to cholera toxin (CT). Immunoblots of pC1 and λB8 lysates probed with anti-CT, exhibited a 30 kDa protein similar to that of pJM17, which carried the ctx gene. Under non-dissociating conditions, anti-CT immunoblots of the same lysates revealed two proteins, one corresponding in size to the holotoxin and the other to CT-A. When analysed by DNA-directed protein synthesis in vitro, both pC1 and λB8 DNA expressed two unique proteins (30 and 11 kDa) similar to that of pJM17.

KW - ELISA

KW - G ganglioside

KW - PGE

KW - Salmonella enterotoxin

KW - cAMP

KW - cholera toxin

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ER -

Biological and immunological characterization of a cloned cholera toxin-like enterotoxin from Salmonella typhimurium

Abstract

Keywords

ASJC Scopus subject areas

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