Synthesis of tetanus toxin fragment C in insect cells by use of a baculovirus expression system.
The baculovirus expression vector p36C was used to express in cells of the insect Spodoptera frugiperda fragment C of tetanus toxin under the control of the strong polyhedrin promoter. Fragment C was expressed intracellularly at a high level and was soluble, allowing it to be purified by affinity chromatography with monoclonal antibody TT08. Purified fragment C from baculovirus was used to immunize mice and was shown to successfully prevent the symptoms of tetanus following a toxin challenge. The ganglioside-binding properties of baculovirus-derived fragment C were compared with those of intact tetanus toxin and native fragment C and were found to be dissimilar.
Charles
I G
IG
Department of Molecular Biology, Wellcome Research Laboratories, Beckenham, Kent, England.
Rodgers
B C
BC
Makoff
A J
AJ
Chatfield
S N
SN
Slater
D E
DE
Fairweather
N F
NF
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D005732:Gangliosides / Q000378:metabolism; D005822:Genetic Vectors; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009036:Moths; D010446:Peptide Fragments / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000378:metabolism; D013744:Tetanus Toxin / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology
D005732:Gangliosides; D010446:Peptide Fragments; D011994:Recombinant Proteins; D013744:Tetanus Toxin; C040635:tetanus toxin fragment C