Summary
A chromosomal DNA segment encoding the biosynthesis of 987P fimbriae was isolated by cosmid-cloning and subsequent subcloning into pBR322. The 12 kb DNA segment expressed five polypeptides with apparent molecular weights of 81,000, 39,000, 28,500, 20,500, and 16,500, respectively. The location of the corresponding genes was determined by insertional mutagenesis using Tn5. The 20.5 K polypeptide was identified as the 987P fimbrial subunit by its reaction with specific anti-987P antibodies. The 81, 39, and 28.5 K polypeptides appeared to be accessory proteins involved in 987P production.
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References
Berg DE, Weiss A, Crossland L (1980) Polarity of Tn5 insertion mutations in Escherichia coli. J Bacteriol 142:439–446
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucl Acids Res 7:1513–1523
Boyer HW, Roulland-Dussoix D (1969) A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol 41:459–472
Brinton CC Jr (1959) Non-flagellar appendages of bacteria. Nature (London) 183:782–786
Dagert M, Ehrlich SD (1979) Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene 6:23–28
Dean EA, Isaacson RE (1982) In vitro adhesion of piliated Escherichia coli to small intestinal villous epithelial cells from rabbits and the identification of a soluble 987P pilus receptor-containing fraction. Infect Immun 36:1192–1198
Dean EA, Isaacson RE (1985) Purification and characterization of a receptor for the 987P pilus of Escherichia coli. Infect Immun 47:98–105
Dougan G, Scherratt D (1977) The transposon Tn1 as a probe for studying ColE2 structure and function. Mol Gen Genet 151:151–160
de Graaf FK, Krenn BE, Klaasen P (1984) Organization and expression of genes involved in the biosynthesis of K99 fimbriae. Infect Immun 43:508–514
Eisenstein BI (1981) Phase variation of type 1 fimbriae in Escherichia coli is under transcriptional control. Science 214:337–339
Grosveld FG, Dahl H-HM, de Boer E, Flavell RA (1981) Isolation of β-globulin-related genes from a human cosmid library. Gene 13:227–237
Isaacson RE, Richter P (1981) Escherichia coli 987P pilus: purification and partial characterization. J Bacteriol 146:784–789
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227:680–685
Mooi FR, de Graaf FK (1985) Molecular biology of fimbriae of enterotoxigenic Escherichia coli. Curr Topics Microbiol Immunol 118:119–138
Mooi FR, de Graaf FK, van Embden JDA (1979) Cloning, mapping and expression of the genetic determinant that encoded for the K88ab antigen. Nucl Acids Res 6:849–865
Mooi FR, Harms N, Bakker D, de Graaf FK (1981) Organization and expression of genes involved in the production of the K88ab antigen. Infect Immun 32:1155–1163
Mooi FR, van Buuren M, Koopman G, Roosendaal B, de Graaf FK (1984) K88ab gene of Escherichia coli encodes a fimbria-like protein distinct from the K88ab fimbrial adhesin. J Bacteriol 159:482–487
Mooi FR, Wijfjes A, de Graaf FK (1983) Identification and characterization of precursors in the biosynthesis of the K88ab fimbria of Escherichia coli. J Bacteriol 154:41–49
Mooi FR, Wouters C, Wijfjes A, de Graaf FK (1982) Construction and characterization of mutants impaired in the biosynthesis of the K88ab antigen. J Bacteriol 150:512–521
Nagy B, Moon HW, Isaacson RE (1976) Colonization of porcine small intestine by Escherichia coli: ileal colonization and adhesion by pig enteropathogens that lack K88 antigen and by some acapsular mutants. Infect Immun 13:1214–1220
Nagy B, Moon HW, Isaacson RE (1977) Colonization of porcine intestine by enterotoxigenic Escherichia coli: selection of piliated forms in vivo, adhesion of piliated forms to epithelial cells in vitro, and incidence of a pilus antigen among porcine enteropathogenic E. coli. Infect Immun 16:344–352
Normark S, Lark D, Hull R, Norgren M, Baga M, O'Hanley P, Schoolnik G, Falkow S (1983) Genetics of digalactoside-binding adhesin from a uropathogenic Escherichia coli strain. Infect Immun 41:942–949
Orndorff PE, Falkow S (1984) Organization and expression of genes responsible for type 1 piliation in Escherichia coli. J Bacteriol 159:736–744
Rhen M, Mäkelä PH, Korhonen TK (1983) P-fimbriae of Escherichia coli are subject to phase variation. FEMS Microbiol Lett 19:267–271
van Embden JDA, van der Donk HJ, van Eijk RV, van der Heide HG, de Jong JA, van Olderen MF, Osterhaus AD, Schouls LM (1983) Molecular cloning and expression of Treponema pallidum DNA in Escherichia coli K-12. Infect Immun 42:187–196
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Communicated by J.W. Lengeler
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de Graaf, F.K., Klaasen, P. Organization and expression of genes involved in the biosynthesis of 987P fimbriae. Molec Gen Genet 204, 75–81 (1986). https://doi.org/10.1007/BF00330190
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DOI: https://doi.org/10.1007/BF00330190