Cloning of the gene coding for Staphylococcus intermedius exfoliative toxin and its expression in Escherichia coli

Terauchi, Rie; Sato, Hisaaki; Endo, You; Aizawa, Chikara; Maehara, Nobutoshi

doi:10.1016/s0378-1135(03)00047-6

Cited by 21 publications

(26 citation statements)

References 34 publications

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“…SIET, the exfoliative toxin from S. intermedius was described by Yamaguchi et al (34), and Terauchi et al (29). The nucleotide sequence of the SIET encoding gene consists of a coding region of 990 bp (30). Our partial sequence showed 100% homology with this gene.…”

Section: Discussionsupporting

confidence: 58%

“…Furthermore, SIET causes exfoliation in 1-day-old chickens, suckling hamsters, and 3-week-old dogs, in contrast to the respective effects of ETs and SHETs (29). The siet gene, located on the chromosomal DNA, consists of a coding region of 990 bp specifying a polypeptide of 330 amino acid residues (30). Until recently, bioassays have been used to detect SIET, while molecular approaches have been searched to detect genes encoding exfoliative toxins (6).…”

Section: Introductionmentioning

confidence: 99%

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Molecular detection of exfoliative toxin in Staphylococcus intermedius isolates from dogs with pyoderma

MÜŞTAK¹

2013

Ankara Üniversitesi Veteriner Fakültesi Dergisi

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Summary:Staphylococcus aureus and S. intermedius are considered as the most significant species causing skin infections in dogs. The aim of this study was to develop a Polymerase Chain Reaction technique for the detection of siet (S. intermedius exfoliative toxin) gene encoding exfoliative toxin in S. intermedius and to investigate its presence in S. intermedius isolates from dogs with pyoderma. A total of 41 isolates (35 S. intermedius, 4 S. aureus, one S. capitis subsp. ureolytica and one S. chromogenes) from dogs with pyoderma were included in the study. Original primers specifically amplifying 145 bp of siet gene and 182 bp of agr (accessory gene regulator) gene locus of S. intermedius were designed in the study. agr gene was detected in all S. intermedius isolates, but not in other isolates. siet gene was detected in all S. intermedius isolates. siet Polymerase Chain Reaction assay was found to be specific since no amplifications were observed with siet negative S. intermedius and other bacterial control strains. Rapid and reliable detection of staphylococci causing skin lesions in dogs and their virulence markers like siet gene will provide important data for clinical practice to manage the disease more effectively by means of treatment and prevention.Key words: Canine pyoderma, exfoliative toxin, siet gene, Staphylococcus intermedius. Piyodermalı köpeklerden izole edilen Staphylococcus intermedius'larda eksfoliatif toksin varlığının moleküler olarak belirlenmesi

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Section: Discussionsupporting

confidence: 58%

Section: Introductionmentioning

confidence: 99%

Molecular detection of exfoliative toxin in Staphylococcus intermedius isolates from dogs with pyoderma

MÜŞTAK¹

2013

Ankara Üniversitesi Veteriner Fakültesi Dergisi

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“…Recently, the cloning of an exfoliative toxin gene from S. intermedius (SIET) was published (29). This toxin, like SHETA from S. hyicus and ETC from S. aureus, is quite different from the serine protease-like group of exfoliative toxins.…”

Section: Discussionmentioning

confidence: 99%

Cloning and Sequence Analysis of Genes Encoding Staphylococcus hyicu s Exfoliative Toxin Types A, B, C, and D

Ahrens

Andresen

2004

J Bacteriol

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Exfoliative toxins produced by certain strains of Staphylococcus hyicus mediate exudative epidermitis in pigs. In this study the genes coding for four different exfoliative toxin from S. hyicus (ExhA, ExhB, ExhC, and ExhD) were cloned and sequenced. The coding sequence of the four toxin genes ranged from 816 to 834 bp. The amino acid sequences of these four toxins were homologous to the earlier described exfoliative toxins SHETB from S. hyicus and ETA, ETB, and ETD from Staphylococcus aureus. The homology between the S. hyicus toxins was at the same level as the homology to the exfoliative toxins from S. aureus. The toxins showed similarity to serine proteases, including preservation of the catalytic tract in ExhA, ExhB, and ExhC. However, in ExhD, Asp in the putative catalytic tract was replaced with Glu. The recombinant toxins could be expressed in Escherichia coli, and three of the four toxins were recognized by monoclonal antibodies raised against native exfoliative toxins.Certain strains of Staphylococcus hyicus can cause the skin disease exudative epidermitis in pigs. Exudative epidermitis is characterized by separation of the cells in the epidermis in the upper stratum spinosum, exfoliation of the skin, erythema, and serous exudation (4, 32). Spontaneous exudative epidermitis in pigs is a generalized infection of the skin caused by strains of S. hyicus, which are able to produce a disease-causing factor designated exfoliative toxin (28). The exfoliative toxins responsible for the characteristic lesion of exudative epidermitis have been identified and purified from strains in Japan and Denmark. These toxins have been characterized as exoproteins of approximately 27 kDa or 30 kDa (6, 24). The toxins isolated in Japan were designated SHETA and SHETB (25), and the toxins isolated in Denmark were designated ExhA, ExhB, and ExhC. One additional toxin antigenically distinct from these was provisionally designated ExhD (5). The toxins differ in their antigenic properties (5, 26); however, no comparison of the toxins isolated in Japan and in Denmark has been published.The complex of skin lesions called the staphylococcal scalded skin syndrome in humans (7, 10, 16) has several aspects in common with exudative epidermitis in pigs. Staphylococcal scalded skin syndrome is caused by infection with Staphylococcus aureus strains that produce exfoliative toxin ETA, ETB, or both (14). The exfoliative toxins from S. aureus, ETA and ETB, have been cloned and their nucleotide sequences have been determined (12,17,20,22). Investigations have shown that the gene encoding ETA is located on the chromosome of S. aureus, while the gene encoding ETB is located on a 42-kb plasmid (reviwed by Arbuthnott [7]). The molecular masses of mature ETA and ETB as deduced from the amino acid sequences were 26,951 Da and 27,318 Da, respectively (17). The differences in the composition of the amino acid sequences reflected the previously observed antigenic differences of the two toxins (8,13,34). A third exfoliative toxin from S. aureus, designat...

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“…In addition, the nucleotide sequence of an S. chromogenes ET is almost identical to that of S. hyicus ExhB, with the replacement of only two amino acids [43]. Conversely, the nucleotide sequences of ETC, SHETA, and SIET are not homologous to those of other ETs and Exhs, and their predicted primary structures do not contain a catalytic triad [5,31,44]. The nucleotides encoding ETC are currently annotated as an enzyme belonging to a group of adenylosuccinate lyases, based on amino acid homologies (GenPept accession No.…”

Section: Open Questionsmentioning

confidence: 99%

“…In addition, two S. hyicus ETs, SHETA and SHETB, and two S. chromogenes ETs were obtained from pigs with exudative epidermitis [29,[41][42][43], and the S. intermedius ET SIETwas isolated from dogs with pyoderma [44]. DNA encoding ETC, SHETA, SHETB, SIET, and an S. chromogenes ET were cloned, and the nucleotide sequences deposited in GenBank [5,31,[43][44][45]. SHETB shares significant amino acid homology with S. aureus ETD [5,45].…”

Section: Open Questionsmentioning

confidence: 99%

Staphylococcal exfoliative toxins: “Molecular scissors” of bacteria that attack the cutaneous defense barrier in mammals

Nishifuji

Sugai

Amagai

2008

Journal of Dermatological Science

138

107

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Cloning of the gene coding for Staphylococcus intermedius exfoliative toxin and its expression in Escherichia coli

Cited by 21 publications

References 34 publications

Molecular detection of exfoliative toxin in Staphylococcus intermedius isolates from dogs with pyoderma

Molecular detection of exfoliative toxin in Staphylococcus intermedius isolates from dogs with pyoderma

Cloning and Sequence Analysis of Genes Encoding Staphylococcus hyicu s Exfoliative Toxin Types A, B, C, and D

Staphylococcal exfoliative toxins: “Molecular scissors” of bacteria that attack the cutaneous defense barrier in mammals

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