Immunogenicity of recombinant F4 （K88） fimbrial adhesin FaeG expressed in tobacco chloroplast  

作　　者：Huifeng Shen Bingjun Qian Weiwei Chen Zhenhua Liu Litao Yang Dabing Zhang Wanqi Liang 

机构地区：[1]School of life Sciences and Biotechnology, Shanghai Jiao tong University, Shanghai 200240, China [2]Bio-X Research Center, Key Laboratory of Genetics & Development and Neuropsychiatric Diseases, Ministry of Education, Shanghai Jiao tong University, Shanghai 200240, China [3]Department of Food Science and Engineering, School of Agriculture and Biology, Shanghai Jiao tong University, Shanghai 200240, China

出　　处：《Acta Biochimica et Biophysica Sinica》2010年第8期558-567,共10页生物化学与生物物理学报（英文版）

摘　　要：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 coil （ETEC） in vivo. This study provides a new alternative for producing the ETEC vaccine using the chloroplast expression system.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 coil （ETEC） in vivo. This study provides a new alternative for producing the ETEC vaccine using the chloroplast expression system.

关 键 词：F4ac enterotoxigenic Escherichia coli FaeG ADHESIN transplastomic tobacco IMMUNOGENICITY 

分 类 号：S858.315.3[农业科学—临床兽医学] Q943.2[农业科学—兽医学]