Immunogenicity of recombinant F4 (K88) fimbrial adhesin FaeG expressed in tobacco chloroplast.
To test the possibility of producing the novel vaccine in plants against diarrhea normally found in neonatal and newly weaned piglets, the faeG gene, encoding a major F4ac fimbrial subunit protein, was introduced into the tobacco chloroplast genome. After two rounds of selection under spectinomycin, we obtained the transgenic plants nearly homoplasmic. RNA gel blot analysis indicated that faeG and the antibiotic selective gene aminoglycoside 3' adenylyltransferase (aadA) were highly transcribed as a dicistron, while the translational level of recombinant FaeG in transplastomic tobacco was about 0.15% of total soluble protein. The immunogenicity of recombinant FaeG produced in tobacco chloroplasts was confirmed by the observation that FaeG-specific antibodies were elicited in mice immunized with total soluble protein of transgenic plants, as well as the result that mouse sera stimulated by chloroplast-derived recombinant FaeG could neutralize F4ac enterotoxigenic Escherichia coli (ETEC) in vivo. This study provides a new alternative for producing the ETEC vaccine using the chloroplast expression system.
Shen
Huifeng
H
School of life Sciences and Biotechnology, Shanghai Jiao tong University, China.
Qian
Bingjun
B
Chen
Weiwei
W
Liu
Zhenhua
Z
Yang
Litao
L
Zhang
Dabing
D
Liang
Wanqi
W
D018830:Adhesins, Escherichia coli / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology; D015152:Blotting, Northern; D015153:Blotting, Western; D002736:Chloroplasts / Q000378:metabolism*; D018742:DNA, Chloroplast / Q000235:genetics; D054307:Enterotoxigenic Escherichia coli / Q000276:immunology; D004927:Escherichia coli Infections / Q000097:blood / Q000276:immunology* / Q000382:microbiology; D022361:Escherichia coli Vaccines / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D009713:Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism
D018830:Adhesins, Escherichia coli; D000907:Antibodies, Bacterial; D018742:DNA, Chloroplast; D022361:Escherichia coli Vaccines; C494388:FaeG protein, E coli; D009713:Nucleotidyltransferases; C022678:streptomycin 3''-adenylyltransferase