Enterohemolysin production is associated with a temperate bacteriophage in Escherichia coli serogroup O26 strains.

Abstract:

:A temperate bacteriophage that determines the expression of enterohemolysin was isolated from Escherichia coli O26 strain C3888. The genetic determinant associated with enterohemolysin production (E-Hly determinant) was cloned from EcoRI-digested bacteriophage DNA in vector plasmid pUC8. pUC8 recombinant plasmid pEO19 carries a 3.7-kb EcoRI insert of phage DNA, and enterohemolysin was expressed in E. coli K-12 after transformation. Hemolysin-negative derivatives of pEO19 were generated by transposon mutagenesis with Tn1725. By subcloning, the phage E-Hly determinant was assigned to a 2,150-bp piece of DNA which is flanked by EcoRI and AccI restriction sites. The enterohemolysin-producing recombinant strains and wild-type strain C3888 express a 60-kDa protein which was detected in the bacterial outer membrane by Western immunoblotting. Biologically active enterohemolysin was detected only in bacteria grown to the stationary phase, and the hemolysin was not released into the culture medium. Lysis of erythrocytes was inhibited by 30 mM dextran 4, which functions as an osmotic protectant without destroying the enterohemolysin itself.

journal_name

J Bacteriol

journal_title

Journal of bacteriology

authors

Beutin L,Bode L,Ozel M,Stephan R

doi

10.1128/jb.172.11.6469-6475.1990

subject

Has Abstract

pub_date

1990-11-01 00:00:00

pages

6469-75

issue

11

eissn

0021-9193

issn

1098-5530

journal_volume

172

pub_type

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