Construction of an epitope vector utilising the diphtheria toxin B-subunit.
An immunogenic loop within the diphtheria toxin has been deleted from the B-subunit by a modification of the inverse polymerase chain reaction (IPCR) and replaced by a unique restriction endonuclease site. An oligonucleotide encoding an identified epitope sequence from the major outer membrane protein of Neisseria meningitidis of similar size and structure to that deleted has been introduced into the restriction site. Expression of the resulting chimeric B-subunit from Escherichia coli yielded a protein that was recognised by a panel of antibodies specific for the meningococcal epitope. Initial immunisation data suggest that this protein could elicit an antibody response against both diphtheria toxin and meningococcal proteins.
Johnson
N
N
University Department of Molecular Microbiology, Southampton General Hospital, UK.
Pickett
M A
MA
Watt
P J
PJ
Clarke
I N
IN
Heckels
J E
JE
D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis; D000942:Antigens, Bacterial / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D003353:Corynebacterium diphtheriae / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D017931:DNA Primers / Q000235:genetics; D004167:Diphtheria Toxin / Q000737:chemistry / Q000235:genetics*; D000939:Epitopes / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D007114:Immunization; D009345:Neisseria meningitidis / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D017931:DNA Primers; D004167:Diphtheria Toxin; D000939:Epitopes; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C039171:diphtheria toxin fragment B