Isolation of a factor causing morphological changes of chinese hamster ovary cells from the culture filtrate of Vibrio parahaemolyticus

Honda, Takeshi; Shimizu, Minako; Takeda, Yoshifumi; Miwatani, Toshio

doi:10.1128/iai.14.4.1028-1033.1976

Cited by 81 publications

(28 citation statements)

References 20 publications

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“…CHO cell assay. The Chinese hamster ovary (CHO) cell assay was carried out as described previously (19). Eagle minimal essential medium was used in place of the F12 medium originally described by Guerrant et al (16) because it decreased the amount of nonspecific elongation of CHO cells seen in the absence of toxin.…”

Section: Fraction Numbermentioning

confidence: 99%

Purification and some properties of a non-o1 Vibrio cholerae enterotoxin that is identical to cholera enterotoxin

Yamamoto¹,

Takeda²,

Miwatani³

et al. 1983

Infect Immun

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Cholera-like enterotoxin was isolated and purified from the culture supernatant of a non-O1 strain of Vibrio cholerae, E8498, isolated from the environment. Enterotoxin was purified by aluminum hydroxide absorption and elution and successive gel filtrations on Sephadex G-100, Bio-Gel A-5m, and Sephadex G-75. Purified enterotoxin gave a single stained band on polyacrylamide gel disc electrophoresis, and the mobility was the same as that of cholera enterotoxin. The specific biological activity of the purified enterotoxin was almost the same as that of cholera enterotoxin in the Chinese hamster ovary cell assay, fluid accumulation in mouse ligated intestine, increase in vascular permeability in rabbit skin, and passive immune hemolysis. Sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis showed that the purified enterotoxin consisted of subunits A and B, identical to those of cholera enterotoxin, and Ouchterlony double gel diffusion tests indicated that the two toxins were immunologically identical. Enterotoxins prepared from several non-O1 strains isolated from human patients were also immunologically identical to cholera enterotoxin.Sodium azide and sodium EDTA were added to culture supernatants at final concentrations of 0.02 and 0.05%, respectively.Purified CT. CT from V. cholerae 569B, serovar 01, purified by the method described by Ohtomo et al. 1128 on July 15, 2020 by guest http://iai.asm.org/ Downloaded from

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Section: Fraction Numbermentioning

confidence: 99%

Purification and some properties of a non-o1 Vibrio cholerae enterotoxin that is identical to cholera enterotoxin

Yamamoto¹,

Takeda²,

Miwatani³

et al. 1983

Infect Immun

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“…Vibrio parahaemolyticus is an enteric pathogen, and shellfish contaminated with this pathogen have been a source of disease outbreaks in Taiwan, Japan, and other coastal regions (5,22,25). A lethal toxin (20,41) and a vascular permeability factor (19), as well as thermostable direct hemolysin (44) and other related hemolysins (18,33,35), have been identified in V. parahaemolyticus. The incidence of V. parahaemolyticus in the environment and shellfish varies greatly, depending on the season, location, sample type, level of fecal pollution, and analytical method (3,8,9,48).…”

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confidence: 99%

Sequence of a cloned pR72H fragment and its use for detection of Vibrio parahaemolyticus in shellfish with the PCR

Lee

Pan

Chen

1995

Appl Environ Microbiol

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The nucleotide sequence of pR72H cloned from Vibrio parahaemolyticus 93 was determined. We examined all V. parahaemolyticus gene sequences published in the GenBank-EMBL databases for homology and found that no other DNA sequence of V. parahaemolyticus was highly homologous to the sequence reported in this study. A pair of primers, VP33-VP32, derived from a pR72H fragment were selected to detect V. parahaemolyticus. The sensitivity of PCR detection for a pure culture of V. parahaemolyticus was 10 cells from crude bacterial lysates. Furthermore, a detection level of 2.6 fg, equivalent to 1 cell, was obtained by using purified chromosomal DNA as the template. The expected PCR products were obtained from all V. parahaemolyticus strains tested (n ‫؍‬ 124), while no PCR amplicons were found in other vibrios or related genera (n ‫؍‬ 50). High levels (10 6 to 10 10 CFU/ml) of Escherichia coli cells did not affect the PCR assay sensitivity. The presence of 10 8 V. parahaemolyticus cells or 10 9 E. coli cells in the PCR mixtures completely inhibited the PCR. When oyster samples were inoculated with V. parahaemolyticus 93 and cultured in tryptic soy broth containing 3% NaCl for 3 h at 35؇C, an initial sample inoculum level of 9.3 CFU/g was detected in a PCR assay with crude bacterial lysates. The PCR assay with enrichment culturing in salt polymyxin broth was compared with the conventional method for naturally contaminated shellfish and fish samples. We conclude that this PCR assay with enrichment culturing is a good alternative method for the detection of V. parahaemolyticus.

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“…The fact that strain K6, a Kp-strain which caused gastroenteritis, lacks the TSD hemolysin gene suggests that some other factor(s) may also be responsible for the gastroenteritis. Incidentally, Honda et al (8) have reported a cholera enterotoxin-like factor in V. parahaemolyticus. The use of these plasmids containing various sizes of WP1 fragments as probes, will offer valuable insight into the pathogenicity of V. parahaemolyticus.…”

Section: Resultsmentioning

confidence: 99%

Cloning and expression in Escherichia coli of Vibrio parahaemolyticus thermostable direct hemolysin and thermolabile hemolysin genes

Taniguchi¹,

Ohta²,

Ogawa³

et al. 1985

J Bacteriol

100

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Two hemolysin genes of Vibrio parahaemolyticus WP1, a thermostable direct (TSD) hemolysin gene and a thermolabile henlolysin gene, were cloned into the pBR322 vector in Escherichia coli K-12 C600. A large amount of the TSD hemolysin produced in E. coli K-12 accumulated in the periplasmic space. The TSD hemolysin gene was localized on a 0.9-kilobase HindIII-BamHI fragment by identifying qualitatively the production of the TSD hemolysin by a reverse passive hemagglutination assay in the osmotic shock fluid. The thermolabile hemolysin gene was isolated on a 1.3-kilobase HindIII-PstI fragment by selection with the hemolysin on blood agar. Southern blot hybridization and colony hybridization experiments indicated that the TSD hemolysin gene was present in the chromosomal DNA of 15 Kanagawa phenomenon-positive strains but not in 14 negative strains, whereas the thermolabile hemolysin gene was detected in all strains. No homologous DNA sequences to TSD and themolabile hemolysin genes were detected in the chromosomes of Vibrio cholerae, Vibrio vulnificus, non-O1 V. cholerae, and Vibrio anguillarum.

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Isolation of a factor causing morphological changes of chinese hamster ovary cells from the culture filtrate of Vibrio parahaemolyticus

Cited by 81 publications

References 20 publications

Purification and some properties of a non-o1 Vibrio cholerae enterotoxin that is identical to cholera enterotoxin

Purification and some properties of a non-o1 Vibrio cholerae enterotoxin that is identical to cholera enterotoxin

Sequence of a cloned pR72H fragment and its use for detection of Vibrio parahaemolyticus in shellfish with the PCR

Cloning and expression in Escherichia coli of Vibrio parahaemolyticus thermostable direct hemolysin and thermolabile hemolysin genes

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