High level production of hybrid potyvirus-like particles carrying repetitive copies of foreign antigens in Escherichia coli.
Synthesis in E. coli of native coat protein of Johnsongrass mosaic virus, and hybrid protein molecules containing foreign antigens, resulted in the intracellular formation of potyvirus-like particles (PVLPs). The foreign antigens used were an octapeptide epitope from Plasmodium falciparum and a decapeptide hormone (luteinizing hormone releasing hormone) at the N- or at both N- and C-terminal regions of the coat protein molecule, and a full length protein antigen (Sj26-glutathione S-transferase of 26 kD from Schistosoma japonicum) replacing the N-terminal 62 amino acids of the coat protein. Electron microscopy of ultrathin sections of E. coli revealed that PVLPs resulting from coat protein molecules containing peptide fusions appeared in vast arrays of parallel strands within the cytoplasm sometimes extending the length of the cell and at times the cells were strung together, with "threads" of PVLPs appearing to connect individual bacterial cells. PVLPs resulting from the fusion of the 26 kD antigen Sj26 to coat protein were shorter and wider. The physical form of the high molecular weight PVLPs enabled purification by simple size exclusion column chromatography. The Sj26-PVLPs administered to mice without adjuvant elicited antibody responses comparable to monomeric Sj26 administered with Freund's Complete Adjuvant.
Jagadish
M N
MN
CSIRO, Division of Biomolecular Engineering, Parkville, Victoria, Australia.
Hamilton
R C
RC
Fernandez
C S
CS
Schoofs
P
P
Davern
K M
KM
Kalnins
H
H
Ward
C W
CW
Nisbet
I T
IT
D000818:Animals; D000941:Antigens / Q000096:biosynthesis* / Q000276:immunology; D000953:Antigens, Protozoan / Q000096:biosynthesis / Q000235:genetics; D001483:Base Sequence; D002213:Capsid / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000378:metabolism*; D005260:Female; D015870:Gene Expression; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000235:genetics; D007987:Gonadotropin-Releasing Hormone / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008811:Mice, Inbred DBA; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009029:Mosaic Viruses; D010963:Plasmodium falciparum / Q000276:immunology; D017800:Potyvirus; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012549:Schistosoma japonicum / Q000201:enzymology; D014771:Virion / Q000276:immunology
D000941:Antigens; D000953:Antigens, Protozoan; D011993:Recombinant Fusion Proteins; D007987:Gonadotropin-Releasing Hormone; D005982:Glutathione Transferase