Abstract
Plasmid pFM205 contains the genetic determinant for the K88ab antigen and is composed of a 4.3-megadalton DNA fragment derived from wild-type K88ab plasmid pRI8801 and cloning vehicle pBR322. The K88 NA of pFM205 contains five genes, which code for polypeptides with apparent molecular weights of 17,000, 26,000 (the K88ab subunit), 27,000 27,500, and 81,000. All five polypeptides were synthesized as precursors approximately 2,000 daltons larger than the mature polypeptides, indicating that they are transported across the cytoplasmic membrane by means of a signal sequence. A set of deletion derivatives of pFM205 was constructed, each containing a deletion in one of the five genes. In strains harboring derivatives of pFM205 containing a deletion in the gene for the 17,000- or 81,000-dalton polypeptide, the K88ab subunit was synthesized and transported to the outside of the cell. However, these strains did not adhere to brushborders or guinea pig erythrocytes, suggesting that the K88ab subunits were not assembled into normal fimbriae. Strains harboring plasmids containing a deletion in the gene for the 27,500-dalton polypeptide still adhered to brush borders and guinea pig erythrocytes, although very little K88ab antigen could be detected with an immunological assay. In strains harboring plasmids containing a deletion in the gene for the 27,000-dalton polypeptide, the K88ab subunit was synthesized but was probably subsequently degraded rapidly.
References
26
Referenced
49
-
Bfrnboim , H. C. , and J. Doly . 1979 . A rapid alkaline extraction procedure for screening recombinant plasmid DNA . Nucleic Acids Res. 7 : 1513 - 1523 .
(
10.1093/nar/7.6.1513
) / Nucleic Acids Res. / A rapid alkaline extraction procedure for screening recombinant plasmid DNA by Bfrnboim H. C. (1979) -
Bolivar , F. , R. L. Rodrlquez , P. J. Green , M. C. Betlach , H. L. Heyneker , H. W. Boyer , J. H. Crosa , and S. Falkow . 1977 . Construction and characterization of new cloning vehicles . Gene 2 : 95 - 113 .
(
10.1016/0378-1119(77)90000-2
) / Gene / Construction and characterization of new cloning vehicles by Bolivar F. (1977) -
Chang , A. C. Y. , and S. N. Cohen . 1978 . Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid . J. Bacteriol. 134 : 1141 - 1156 .
(
10.1128/JB.134.3.1141-1156.1978
) / J. Bacteriol. / Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid by Chang A. C. Y. (1978) -
Dagert , M. , and S. D. Ehrlich . 1979 . Prolonged incubation in calcium chloride improves the competence of Escherichia coli celis . Gene 6 : 23 - 28 .
(
10.1016/0378-1119(79)90082-9
) / Gene / Prolonged incubation in calcium chloride improves the competence of Escherichia coli celis by Dagert M. (1979) -
Davis , D. B. , and P. C. Tal . 1980 . The mechanism of protein secretion across membranes . Nature (London) 283 : 433 - 438 .
(
10.1038/283433a0
) / Nature (London) / The mechanism of protein secretion across membranes by Davis D. B. (1980) -
Dougan , G. , and D. Sherratt . 1977 . The transposon Tnl as a probe for studying ColEl structure and function . Mol. Gen. Genet. 151 : 151 - 160 .
(
10.1007/BF00338689
) / Mol. Gen. Genet. / The transposon Tnl as a probe for studying ColEl structure and function by Dougan G. (1977) -
Gaastra , W. , P. Klemm , J. M. Walker , and F. K. de Graaf . 1979 . K88 fimbrial proteins: amino and carboxyl terminal sequences of intact proteins and cyanogen bromide fragments . FEMS Lett. 6 : 15 - 18 .
(
10.1111/j.1574-6968.1979.tb04267.x
) / FEMS Lett. / K88 fimbrial proteins: amino and carboxyl terminal sequences of intact proteins and cyanogen bromide fragments by Gaastra W. (1979) -
. Gaastra , W. , F. R. Mool , A. R. StultJe , and F. K. de Graaf . 1981 . The nucleotide sequence of the gene encoding the K88ab protein subunit of porcine enterotoxigenic Escherichia coli . FEMS Lett. 12 : 41 - 46 .
(
10.1111/j.1574-6968.1981.tb07608.x
) / FEMS Lett. / The nucleotide sequence of the gene encoding the K88ab protein subunit of porcine enterotoxigenic Escherichia coli by Gaastra W. (1981) -
Goodman , H. M. , and R. J. MacDonald . 1979 . Cloning of hormone genes from a mixture of cDNA molecules . Methods Enzymol. 68 : 75 - 89 .
(
10.1016/0076-6879(79)68007-2
) / Methods Enzymol. / Cloning of hormone genes from a mixture of cDNA molecules by Goodman H. M. (1979) -
Guin 6e, P. A. M. , and W. H. Jansen . 1979 . Behavior of Escherichia coli K antigens K88ab, K88ac, and K88ad in immunoelectrophoresis, double diffusion, and hemagglutination . Infect. Immun. 23 : 700 - 705 .
(
10.1128/IAI.23.3.700-705.1979
) / Infect. Immun. / Behavior of Escherichia coli K antigens K88ab, K88ac, and K88ad in immunoelectrophoresis, double diffusion, and hemagglutination by Guin P. A. M. (1979) - Jones , G. W. , and J. M. Rutter . 1972 . Role of the K88 antigen in the pathogenesis of neonatal diarrhea caused by Escherichia coli in piglets . Infect. Immun. 5 : 595 - 605 . / Infect. Immun. / Role of the K88 antigen in the pathogenesis of neonatal diarrhea caused by Escherichia coli in piglets by Jones G. W. (1972)
-
Kehoe , M. , R. Sellwoud , P. Shipley , and G. Dougan . 1981 . Genetic analysis of K88-mediated adhesion of enterotoxigenic Escherichia coli . Nature (London) 291 : 122 - 126 .
(
10.1038/291122a0
) / Nature (London) / Genetic analysis of K88-mediated adhesion of enterotoxigenic Escherichia coli by Kehoe M. (1981) -
Mool , F. R. , and F. K. de Graaf . 1979 . Isolation and characterization of K88 antigens . FEMS Lett. 5 : 17 - 20 .
(
10.1111/j.1574-6968.1979.tb03237.x
) / FEMS Lett. / Isolation and characterization of K88 antigens by Mool F. R. (1979) -
Mool , F. R. , F. K. de Graaf , and J. D. A. van Embden . 1979 . Cloning, mapping and expression of the genetic determinant that encodes for the K88ab antigen . Nucleic Acids Res. 6 : 849 - 865 .
(
10.1093/nar/6.3.849
) / Nucleic Acids Res. / Cloning, mapping and expression of the genetic determinant that encodes for the K88ab antigen by Mool F. R. (1979) -
Mool , F. R. , N. Harms , D. Bakker , and F. K. de Graaf . 1981 . Organization and expression of genes involved in the production of the K88ab antigen . Infect. Immun. 32 : 1155 - 1163 .
(
10.1128/IAI.32.3.1155-1163.1981
) / Infect. Immun. / Organization and expression of genes involved in the production of the K88ab antigen by Mool F. R. (1981) -
Murray , K. , and N. E. Murray . 1975 . Phage lambda receptor chromosomes for DNA fragments made with HindIII and EcoRI . J. Mol. Biol. 98 : 551 - 564 .
(
10.1016/S0022-2836(75)80086-6
) / J. Mol. Biol. / Phage lambda receptor chromosomes for DNA fragments made with HindIII and EcoRI by Murray K. (1975) - 0rskow, I., F. 0rskov, W. J. Sojka , and W. Wittig. 1964 . K antigens K88ab (L) and K88ac (L) in E. coli . Acta Pathol. Microbiol. Scand. 62 : 439 - 447 . / Acta Pathol. Microbiol. Scand. / K antigens K88ab (L) and K88ac (L) in E. coli by Sojka W. (1964)
-
Palva , E. T. , T. R. Hfrst , S. J. S. Hardy , J. Hohngren , and L. Randall . 1981 . Synthesis of a precursor to the 1B subunit of heat-labile enterotoxin in Escherichia coli . J. Bacteriol. 146 : 325 - 330 .
(
10.1128/jb.146.1.325-330.1981
) / J. Bacteriol. / Synthesis of a precursor to the 1B subunit of heat-labile enterotoxin in Escherichia coli by Palva E. T. (1981) -
Parker , C. M. , R. M. Watson , and J. Vinograd . 1977 . Mapping of closed circular DNA's by cleavage with restriction endonucleases and calibration by agarose gel electrophoresis . Proc. Natl. Acad. Sci. U.S.A. 74 : 851 - 855 .
(
10.1073/pnas.74.3.851
) / Proc. Natl. Acad. Sci. U.S.A. / Mapping of closed circular DNA's by cleavage with restriction endonucleases and calibration by agarose gel electrophoresis by Parker C. M. (1977) -
Rodriquez , R. L. , R. W. West , H. L. Heyneker , F. Bollvar , and H. W. Boyer . 1979 . Characterizing wild-type and mutant promotors of the tetracycline resistance gene in pBR313 . Nucleic Acids Res. 6 : 3267 - 3286 .
(
10.1093/nar/6.10.3267
) / Nucleic Acids Res. / Characterizing wild-type and mutant promotors of the tetracycline resistance gene in pBR313 by Rodriquez R. L. (1979) -
Shipley , P. L. , G. Dougan , and S. F. Falkow . 1981 . Identification and cloning of the genetic determinant that encodes for the K88ac adherence antigen . J. Bacteriol. 145 : 920 - 925 .
(
10.1128/JB.145.2.920-925.1981
) / J. Bacteriol. / Identification and cloning of the genetic determinant that encodes for the K88ac adherence antigen by Shipley P. L. (1981) -
Sttrm , S. , F. Orskov , I. 0rskov, and A. Birch-Andersen . 1967 . Episome-carried surface antigen K88 of Escherichia coli . J. Bacteriol. 93 : 740 - 748 .
(
10.1128/JB.93.2.740-748.1967
) / J. Bacteriol. / Episome-carried surface antigen K88 of Escherichia coli by Sttrm S. (1967) -
Sutclffe , J. G. 1978 . pBR322 restriction map derived from the DNA sequence: accurate DNA size markers up to 4361 nucleotide pairs long . Nucleic Acids Res. 5 : 2721 - 2728 .
(
10.1093/nar/5.8.2721
) / Nucleic Acids Res. / pBR322 restriction map derived from the DNA sequence: accurate DNA size markers up to 4361 nucleotide pairs long by Sutclffe J. G. (1978) -
Thompson , R. , and M. Achtman . 1979 . The control region of the F sex factor DNA transfer cistrons: physical mapping by deletion analysis . Mol. Gen. Genet. 169 : 49 - 57 .
(
10.1007/BF00267544
) / Mol. Gen. Genet. / The control region of the F sex factor DNA transfer cistrons: physical mapping by deletion analysis by Thompson R. (1979) -
. Van Doom , J. , B. Oudega , F. R. Moot , and F. K. de Graaf . 1982 . Subceliular localization of polypeptides involved in the biosynthesis of the K88ab fimbriae . FEMS Lett. 13 : 99 - 104 .
(
10.1111/j.1574-6968.1982.tb08235.x
) / FEMS Lett. / Subceliular localization of polypeptides involved in the biosynthesis of the K88ab fimbriae by Van Doom J. (1982) -
W Uson , M. R. , and A. W. Hohmann . 1974 . Immunity to Escherichia coli in pigs: adhesion of enteropathogenic Escherichia coli to isolated intestinal epithelial cells . Infect. Immun. 10 : 776 - 782 .
(
10.1128/IAI.10.4.776-782.1974
) / Infect. Immun. / Immunity to Escherichia coli in pigs: adhesion of enteropathogenic Escherichia coli to isolated intestinal epithelial cells by Uson M. R. (1974)
Dates
Type | When |
---|---|
Created | 5 years, 8 months ago (Jan. 3, 2020, 12:32 p.m.) |
Deposited | 4 years, 1 month ago (July 29, 2021, 1:27 p.m.) |
Indexed | 1 year, 7 months ago (Jan. 18, 2024, 12:26 a.m.) |
Issued | 43 years, 4 months ago (May 1, 1982) |
Published | 43 years, 4 months ago (May 1, 1982) |
Published Print | 43 years, 4 months ago (May 1, 1982) |
@article{Mooi_1982, title={Construction and characterization of mutants impaired in the biosynthesis of the K88ab antigen}, volume={150}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.150.2.512-521.1982}, DOI={10.1128/jb.150.2.512-521.1982}, number={2}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Mooi, F R and Wouters, C and Wijfjes, A and de Graaf, F K}, year={1982}, month=may, pages={512–521} }