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

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

Research output: Contribution to journalArticle

25 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)315-329
Number of pages15
JournalMicrobial Pathogenesis
Volume9
Issue number5
DOIs
StatePublished - 1990

Fingerprint

Cholera Toxin
Enterotoxins
Salmonella typhimurium
Genes
DNA
Proteins
Escherichia coli
G(M1) Ganglioside
Fluids and Secretions
Oligonucleotide Probes
Cricetulus
Dinoprostone
Bacteriophages
Immune Sera
Ovary
Plasmids
Clone Cells
Rabbits

Keywords

  • cAMP
  • cholera toxin
  • ELISA
  • G ganglioside
  • PGE
  • Salmonella enterotoxin

ASJC Scopus subject areas

  • Microbiology
  • Infectious Diseases

Cite this

Biological and immunological characterization of a cloned cholera toxin-like enterotoxin from Salmonella typhimurium. / Prasad, Rajendra; Chopra, Ashok; Peterson, Johnny; Pericas, Roser; Houston, Clifford W.

In: Microbial Pathogenesis, Vol. 9, No. 5, 1990, p. 315-329.

Research output: Contribution to journalArticle

@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 = "cAMP, cholera toxin, ELISA, G ganglioside, PGE, Salmonella enterotoxin",
author = "Rajendra Prasad and Ashok Chopra and Johnny Peterson and Roser Pericas and Houston, {Clifford W.}",
year = "1990",
doi = "10.1016/0882-4010(90)90066-Y",
language = "English (US)",
volume = "9",
pages = "315--329",
journal = "Microbial Pathogenesis",
issn = "0882-4010",
publisher = "Academic Press Inc.",
number = "5",

}

TY - JOUR

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

AU - Prasad, Rajendra

AU - Chopra, Ashok

AU - Peterson, Johnny

AU - Pericas, Roser

AU - Houston, Clifford W.

PY - 1990

Y1 - 1990

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 - cAMP

KW - cholera toxin

KW - ELISA

KW - G ganglioside

KW - PGE

KW - Salmonella enterotoxin

UR - http://www.scopus.com/inward/record.url?scp=0025688641&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025688641&partnerID=8YFLogxK

U2 - 10.1016/0882-4010(90)90066-Y

DO - 10.1016/0882-4010(90)90066-Y

M3 - Article

C2 - 2099384

AN - SCOPUS:0025688641

VL - 9

SP - 315

EP - 329

JO - Microbial Pathogenesis

JF - Microbial Pathogenesis

SN - 0882-4010

IS - 5

ER -