Abstract
From a cosmid clone, a 5.3-kilobase (kb) HindIII fragment of Salmonella enteritidis DNA containing the fimA gene was subcloned into bacteriophage T7 promoter vectors in both orientations. Predominantly mature fimbrin (14,000 Mr) was produced by clones containing the 5.3-kb insert, whereas the original cosmid clone predominantly accumulated a prefimbrin precursor (16,500 Mr). T7 RNA polymerase-dependent expression of the 5.3-kb insert and of a series of nested deletions revealed several membrane-localized polypeptides (80,000, 40,000, 29,000, 25,000, and 16,500 Mrs) transcribed in the same direction as fimA as well as a single polypeptide (9,000 Mr) transcribed in the opposite direction. Mini-Mu and TnphoA (Tn5 IS50L::phoA) transposon mutagenesis was used to identify a 2- to 3.5-kb region downstream of fimA that affected fimbrin production and processing. A more distant region (greater than 7 kb), revealed by Tn10 and Mu dI mutagenesis, was also required for fimbriation but did not hybridize with the 5.3-kb fragment. Yet another distant region did hybridize to the 5.3-kb fragment, suggesting the existence of other fimbriation-related genes.
MeSH Terms
Bacterial Proteins/genetics,isolation & purification
Blotting, Western
Coliphages/genetics
Escherichia coli/genetics
Fimbriae, Bacterial/physiology
Genes, Bacterial
Genotype
Molecular Weight
Mutation
Plasmids
Restriction Mapping
Salmonella enteritidis/genetics,physiology
Salmonella typhimurium/genetics
Chemicals
Bacterial Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Müller K H
Department of Biochemistry and Microbiology, University of Victoria, British Columbia, Canada.
Trust T J
Kay W W
References (32)
32 references, click to expand
-
An improved DNA sequencing strategy.
Anal Biochem. 1985 May 15;147(1):114-9
PMID: 2992313
-
Single-stranded DNA 'blue' T7 promoter plasmids: a versatile tandem promoter system for cloning and protein engineering.
Protein Eng. 1986 Oct-Nov;1(1):67-74
PMID: 3507689
-
Two modes of control of pilA, the gene encoding type 1 pilin in Escherichia coli.
J Bacteriol. 1985 Oct;164(1):321-30
PMID: 3930469
-
Rapid transfer of DNA from agarose gels to nylon membranes.
Nucleic Acids Res. 1985 Oct 25;13(20):7207-21
PMID: 4059056
-
TnphoA: a transposon probe for protein export signals.
Proc Natl Acad Sci U S A. 1985 Dec;82(23):8129-33
PMID: 2999794
-
Transcriptional activation of a pap pilus virulence operon from uropathogenic Escherichia coli.
EMBO J. 1985 Dec 30;4(13B):3887-93
PMID: 2868893
-
The genetic determinant of adhesive function in type 1 fimbriae of Escherichia coli is distinct from the gene encoding the fimbrial subunit.
J Bacteriol. 1986 Mar;165(3):1033-6
PMID: 2419305
-
Piliation control mechanisms in Neisseria gonorrhoeae.
Proc Natl Acad Sci U S A. 1986 Jun;83(11):3890-4
PMID: 2872674
-
Two regulatory fim genes, fimB and fimE, control the phase variation of type 1 fimbriae in Escherichia coli.
EMBO J. 1986 Jun;5(6):1389-93
PMID: 2874022
-
Purification and characterization of fimbriae from Salmonella enteritidis.
J Bacteriol. 1986 Oct;168(1):221-7
PMID: 2875990
-
Gene conversion involving the pilin structural gene correlates with pilus+ in equilibrium with pilus- changes in Neisseria gonorrhoeae.
Cell. 1986 Oct 24;47(2):267-76
PMID: 2876777
-
Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes.
J Mol Biol. 1986 May 5;189(1):113-30
PMID: 3537305
-
Characterization of genes encoding type 1 fimbriae of Klebsiella pneumoniae, Salmonella typhimurium, and Serratia marcescens.
Infect Immun. 1987 Feb;55(2):281-7
PMID: 2879791
-
Identification and characterization of genes determining receptor binding and pilus length of Escherichia coli type 1 pili.
J Bacteriol. 1987 Feb;169(2):640-5
PMID: 2879830
-
Hemagglutination of human group A erythrocytes by enterotoxigenic Escherichia coli isolated from adults with diarrhea: correlation with colonization factor.
Infect Immun. 1977 Nov;18(2):330-7
PMID: 336541
-
Prevention of colonization of the urinary tract of mice with Escherichia coli by blocking of bacterial adherence with methyl alpha-D-mannopyranoside.
J Infect Dis. 1979 Mar;139(3):329-32
PMID: 376757
-
A small cosmid for efficient cloning of large DNA fragments.
Gene. 1980 Nov;11(3-4):291-8
PMID: 6260575
-
Host-specific fimbrial adhesins of noninvasive enterotoxigenic Escherichia coli strains.
Microbiol Rev. 1982 Jun;46(2):129-61
PMID: 6126799
-
A prokaryotic membrane anchor sequence: carboxyl terminus of bacteriophage f1 gene III protein retains it in the membrane.
Proc Natl Acad Sci U S A. 1982 Sep;79(17):5200-4
PMID: 6291030
-
Plasmid insertion mutagenesis and lac gene fusion with mini-mu bacteriophage transposons.
J Bacteriol. 1984 May;158(2):488-95
PMID: 6327606
-
Genes of pyelonephritogenic E. coli required for digalactoside-specific agglutination of human cells.
EMBO J. 1984 May;3(5):1167-73
PMID: 6145590
-
Organization and expression of genes responsible for type 1 piliation in Escherichia coli.
J Bacteriol. 1984 Aug;159(2):736-44
PMID: 6146599
-
Reduction of the minimal sequence for initiation of DNA synthesis by qualitative or quantitative changes of an initiator protein.
Nature. 1984 Sep 20-26;311(5983):279-80
PMID: 6541296
-
A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes.
Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074-8
PMID: 3156376
-
Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors.
Gene. 1985;33(1):103-19
PMID: 2985470
-
Enterobacterial fimbriae.
J Bacteriol. 1987 Mar;169(3):934-8
PMID: 2880835
-
Localization of the receptor-binding protein adhesin at the tip of the bacterial pilus.
Nature. 1987 Jul 2-8;328(6125):84-7
PMID: 2885755
-
Fimbrial phase variation in Escherichia coli: dependence on integration host factor and homologies with other site-specific recombinases.
J Bacteriol. 1987 Aug;169(8):3840-3
PMID: 2886490
-
Integration host factor is required for the DNA inversion that controls phase variation in Escherichia coli.
Proc Natl Acad Sci U S A. 1987 Sep;84(18):6506-10
PMID: 2888114
-
Vectors for selective expression of cloned DNAs by T7 RNA polymerase.
Gene. 1987;56(1):125-35
PMID: 3315856
-
Cloning and expression of a Salmonella enteritidis fimbrin gene in Escherichia coli.
J Bacteriol. 1988 Sep;170(9):4216-22
PMID: 2900832
-
An invertible element of DNA controls phase variation of type 1 fimbriae of Escherichia coli.
Proc Natl Acad Sci U S A. 1985 Sep;82(17):5724-7
PMID: 2863818