Cloning and characterization of the hemolysin determinants from Vibrio cholerae RV79(Hly+), RV79(Hly-), and 569B

The Hly region from the chromosome of Vibrio cholerae El Tor strain RV79(Hly-) and the nonhemolytic classical strain 569B were cloned into plasmid vector pBR322. Escherichia coli K-12 transformants possessing these recombinant plasmids were nonhemolytic and were detected with a 32P-labeled hly-specific DNA probe. Restriction endonuclease Sau3AI digestions of the cloned hly loci of two independently obtained RV79(Hly+) convertants, when compared with the digests of cloned RV79(Hly-) loci, revealed that an apparent alteration (10 to 15 base pairs) had occurred. In contrast, an apparent 20-base-pair deletion was present in the cloned hly locus of the classical biotype V. cholerae strain 569B. Maxicell analysis and immunoprecipitation of labeled proteins of E. coli which are encoded by the cloned hly loci of RV79(Hly+) and from nuclease BAL 31-deleted plasmids, as well as immunoprecipitation of [35S]methionine-labeled V. cholerae proteins, suggest that the hemolysin is an 84,000-dalton polypeptide.

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

[2]  J. Murphy,et al.  Molecular cloning of the hemolysin determinant from Vibrio cholerae El Tor , 1984, Journal of bacteriology.

[3]  S. Jones,et al.  Small DNA deletions creating avirulence in Streptococcus pyogenes. , 1984, Science.

[4]  Y. Takeda,et al.  Non-O1 Vibrio cholerae hemolysin: purification, partial characterization, and immunological relatedness to El Tor hemolysin , 1984, Infection and immunity.

[5]  N. Fairweather,et al.  Expression of a Cloned Staphylococcus aureus α-Hemolysin Determinant in Bacillus subtilis and Staphylococcus aureus , 1984 .

[6]  T. Meyer,et al.  Pilus expression in neisseria gonorrhoeae involves chromosomal rearrangement , 1982, Cell.

[7]  S. Moseley,et al.  Nucleotide sequence homology between the heat-labile enterotoxin gene of Escherichia coli and Vibrio cholerae deoxyribonucleic acid , 1980, Journal of bacteriology.

[8]  T. Honda,et al.  Purification and characterization of a hemolysin produced by Vibrio cholerae biotype El Tor: another toxic substance produced by cholera vibrios , 1979, Infection and immunity.

[9]  A. Sancar,et al.  Simple method for identification of plasmid-coded proteins , 1979, Journal of bacteriology.

[10]  M. Simon,et al.  Recombinational switch for gene expression. , 1977, Science.

[11]  D. Hogness,et al.  Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[12]  A. H. Al-dujaili,et al.  Pseudomonas aeruginosa infection in hospital: a comparison between ‘infective’ and ‘environmental’ strains , 1975, Journal of Hygiene.

[13]  S. H. Richardson,et al.  Biochemistry of Vibrio cholerae Virulence III. Nutritional Requirements for Toxin Production and the Effects of pH on Toxin Elaboration in Chemically Defined Media , 1973, Infection and immunity.

[14]  M Mandel,et al.  Calcium-dependent bacteriophage DNA infection. , 1970, Journal of molecular biology.

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

[16]  W. R. Romig,et al.  Production of Bacteriophage-Associated Materials by Vibrio cholerae: Possible Correlation with Pathogenicity , 1970, Infection and immunity.

[17]  S. Falkow,et al.  Polynucleotide Sequence Relationships among Members of Enterobacteriaceae , 1969, Journal of bacteriology.

[18]  S. Falkow,et al.  Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. , 1977, Gene.