Cloning of the fomA gene, encoding the major outer membrane porin of Fusobacterium nucleatum ATCC10953

TY - JOUR

T1 - Cloning of the fomA gene, encoding the major outer membrane porin of Fusobacterium nucleatum ATCC10953

AU - Jensen, H B

AU - Skeidsvoll, J

AU - Fjellbirkeland, A

AU - Høgh, B

AU - Puntervoll, P

AU - Kleivdal, H

AU - Tommassen, J

PY - 1996/11

Y1 - 1996/11

N2 - The major outer membrane protein, FomA, of the Gram-negative human oral pathogen Fusobacterium nucleatum functions as a porin and is assumed to act as a receptor protein in coaggregation with other oral pathogenic bacteria such as Streptococcus sanguis and Porphyromonas gingivalis. We describe here the cloning of fomA from F. nucleatum in E. coli. Using pGEM3Zf(+), three recombinant plasmids were carrying parts of the fomA gene, but none of these contained regions upstream of the coding sequence. From these plasmids a clone was constructed which contained the whole fomA gene. The ATCC 10953 fomA gene was cloned under the phosphate limitation-inducible phoE promoter, using a vector derived from pACYC184. The protein was found to be incorporated into the outer membrane of the host in an apparently normal manner, as judged by heat-modifiability, trypsin-accessibility, and accessibility to antibodies to the protein in a whole cell enzyme-linked immunosorbent assay. The cloned FomA was found to exhibit pore-forming activity.

AB - The major outer membrane protein, FomA, of the Gram-negative human oral pathogen Fusobacterium nucleatum functions as a porin and is assumed to act as a receptor protein in coaggregation with other oral pathogenic bacteria such as Streptococcus sanguis and Porphyromonas gingivalis. We describe here the cloning of fomA from F. nucleatum in E. coli. Using pGEM3Zf(+), three recombinant plasmids were carrying parts of the fomA gene, but none of these contained regions upstream of the coding sequence. From these plasmids a clone was constructed which contained the whole fomA gene. The ATCC 10953 fomA gene was cloned under the phosphate limitation-inducible phoE promoter, using a vector derived from pACYC184. The protein was found to be incorporated into the outer membrane of the host in an apparently normal manner, as judged by heat-modifiability, trypsin-accessibility, and accessibility to antibodies to the protein in a whole cell enzyme-linked immunosorbent assay. The cloned FomA was found to exhibit pore-forming activity.

KW - Bacterial Outer Membrane Proteins/genetics

KW - Cell Membrane

KW - Cloning, Molecular

KW - Fusobacterium nucleatum/genetics

KW - Gene Expression

KW - Genes, Bacterial

KW - Humans

KW - Phosphates

KW - Porins/genetics

KW - Promoter Regions, Genetic

U2 - 10.1006/mpat.1996.0066

DO - 10.1006/mpat.1996.0066

M3 - Journal article

C2 - 8938641

SN - 0882-4010

VL - 21

SP - 331

EP - 342

JO - Microbial Pathogenesis

JF - Microbial Pathogenesis

IS - 5

ER -