Cloning of genes for production of Escherichia coli Shiga-like toxin type II

Genes controlling production of Shiga-like toxin type II (SLT-II) in Escherichia coli were cloned from the SLT-II-converting bacteriophage 933W and compared with the Shiga-like toxin type I (SLT-I) genes previously isolated and described from phage 933J. Subcloning analysis identified a region within the 4.9-kilobase EcoRI fragment of phage 933W that was associated with SLT-II production. Experiments with E. coli minicells containing these subclones demonstrated that the 4.9-kilobase EcoRI fragment encodes the structural genes for SLT-II. These experiments additionally showed the genetic organization of the SLT-II genes to be the same as that of the SLT-I genes, with the coding sequence for the large A subunit adjacent to that for the smaller B subunit. The mobilities of the SLT-II subunits in sodium dodecyl sulfate-polyacrylamide gels were slightly greater than those determined for the SLT-I subunits. Although apparent processing of the SLT-I subunits was observed with polymyxin B treatment of the labeled minicells, no processing of the SLT-II subunits was detected. Southern blot hybridization studies suggested that the DNA fragment carrying the SLT-II structural genes shares approximately 50 to 60% homology with the DNA of the SLT-I structural genes.

[1]  B. Rowe,et al.  Heterogeneity of Escherichia coli phages encoding Vero cytotoxins: comparison of cloned sequences determining VT1 and VT2 and development of specific gene probes. , 1987, Journal of general microbiology.

[2]  M. P. Jackson,et al.  Nucleotide sequence analysis of the structural genes for Shiga-like toxin I encoded by bacteriophage 933J from Escherichia coli. , 1987, Microbial pathogenesis.

[3]  J. Wells,et al.  Production of Shiga-like toxin by Escherichia coli. , 1986, The Journal of infectious diseases.

[4]  R. Holmes,et al.  Two toxin-converting phages from Escherichia coli O157:H7 strain 933 encode antigenically distinct toxins with similar biologic activities , 1986, Infection and immunity.

[5]  J. Friesen,et al.  Cloning and expression of the genes specifying Shiga-like toxin production in Escherichia coli H19 , 1986, Journal of bacteriology.

[6]  B. Rowe,et al.  Cloning of genes determining the production of vero cytotoxin by Escherichia coli. , 1985, Journal of general microbiology.

[7]  R. Holmes,et al.  Cloning of Shiga-like toxin structural genes from a toxin converting phage of Escherichia coli. , 1985, Science.

[8]  L. Baron,et al.  Development of a DNA probe to detect Salmonella typhi , 1985, Journal of clinical microbiology.

[9]  J. Cantey.  Shiga toxin--an expanding role in the pathogenesis of infectious diseases. , 1985, The Journal of infectious diseases.

[10]  R. Holmes,et al.  Cloning of extracellular DNase and construction of a DNase-negative strain of Vibrio cholerae , 1985, Infection and immunity.

[11]  L. Pickering,et al.  Shiga-like cytotoxin production by enteropathogenic Escherichia coli serogroups , 1985, Infection and immunity.

[12]  R. Holmes,et al.  Shiga-like toxin-converting phages from Escherichia coli strains that cause hemorrhagic colitis or infantile diarrhea. , 1984, Science.

[13]  L. Baron,et al.  Expression of lipopolysaccharide O antigen in Escherichia coli K-12 hybrids containing plasmid and chromosomal genes from Shigella dysenteriae 1 , 1984, Infection and immunity.

[14]  Guidelines for research involving recombinant DNA molecules. , 1984, Federal register.

[15]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[16]  J. Messing,et al.  Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. , 1983, Gene.

[17]  H. Smith,et al.  Vero cell toxins in Escherichia coli and related bacteria: transfer by phage and conjugation and toxic action in laboratory animals, chickens and pigs. , 1983, Journal of general microbiology.

[18]  A. O’Brien,et al.  Purification and characterization of a Shigella dysenteriae 1-like toxin produced by Escherichia coli , 1983, Infection and immunity.

[19]  D. Griffin,et al.  Release of Shiga Toxin from Shigella dysenteriae 1 by Polymyxin B , 1983, Infection and immunity.

[20]  M. Thompson,et al.  Production of Shigella dysenteriae type 1-like cytotoxin by Escherichia coli. , 1982, The Journal of infectious diseases.

[21]  A. O’Brien,et al.  Immunochemical and cytotoxic activities of Shigella dysenteriae 1 (shiga) and shiga-like toxins , 1982, Infection and immunity.

[22]  S. Olsnes,et al.  Subunit structure of Shigella cytotoxin. , 1981, The Journal of biological chemistry.

[23]  D. Griffin,et al.  Characterization of Shigella dysenteriae 1 (Shiga) toxin purified by anti-Shiga toxin affinity chromatography , 1980, Infection and immunity.

[24]  J. Dalrymple,et al.  Quantitative microtiter cytotoxicity assay for Shigella toxin , 1980, Journal of clinical microbiology.

[25]  D. Botstein,et al.  Advanced Bacterial Genetics: A Manual for Genetic Engineering , 1980 .

[26]  F. Bolivar,et al.  Construction and characterization of new cloning vehicles. IV. Deletion derivatives of pBR322 and pBR325. , 1980, Gene.

[27]  H. Birnboim,et al.  A rapid alkaline extraction procedure for screening recombinant plasmid DNA. , 1979, Nucleic acids research.

[28]  N. Tolbert,et al.  A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. , 1978, Analytical biochemistry.

[29]  P. Gemski,et al.  Cellular release of heat-labile enterotoxin of Escherichia coli by bacteriophage induction , 1978, Infection and immunity.

[30]  J Konowalchuk,et al.  Vero response to a cytotoxin of Escherichia coli , 1977, Infection and immunity.

[31]  E. Southern.  Detection of specific sequences among DNA fragments separated by gel electrophoresis. , 1975, Journal of molecular biology.

[32]  R. Curtiss,et al.  Production, properties and utility of bacterial minicells. , 1975, Current topics in microbiology and immunology.

[33]  L. Hsu,et al.  Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. , 1972, Proceedings of the National Academy of Sciences of the United States of America.

[34]  Jeffrey H. Miller.  Experiments in molecular genetics , 1972 .

[35]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[36]  H. Boyer,et al.  A complementation analysis of the restriction and modification of DNA in Escherichia coli. , 1969, Journal of molecular biology.

[37]  R. Appleyard.  Segregation of New Lysogenic Types during Growth of a Doubly Lysogenic Strain Derived from Escherichia Coli K12. , 1954, Genetics.