Expression of the Antigen from Bordetella pertussis in Transgenic Tobacco and its Oral Immunogenicity in Mice

• Main Authors: Meng-Huei Chang, 張萌惠
• Other Authors: Shih-Tong Jeng
• Format: Others
• Language: zh-TW
• Published: 2001
• Online Access: http://ndltd.ncl.edu.tw/handle/48185625742274623799

碩士 === 國立臺灣大學 === 植物學研究所 === 89 === Whooping cough is a disease caused by Bordetella pertussis. Following infection, the virulent bacteria attach to the ciliated epithelial cells of the respiratory tract, and cause acute cough called whooping cough. The heat-killed bacteria are the major component of whole-cell vaccines used currently, but it causes different levels of side effect including life threatened. It is critical to design acellular vaccines for safety. The virulent factors such as filamentous hemagglutinin and pertussis toxin are good candidates for vaccines of whooping cough. Plants can be transformed to produce transgenic protein and cultivated on large scale easily. Transgenic tobacco and tomato were made with the gene encoding the Fragment A of filamentous hemagglutinin. Fragment A was modified by adding amino acid sequence GSGC and ER retention signal HDEL at the C terminus for facilitating protein in dimer form and aggregating in ER. Transgenic tobacco was also made with the gene encoding pertussis toxin S1 subunit and the gene was modified by adding HDEL at C terminus. Transformations were finished by Agrobacteria, and transgenic plants were selected by genomic DNA PCR. The transcription of transgenes was detected by RT-PCR, and the expression of protein was analyzed by western blotting. Also, the antigen expressed by E. coli was used to evaluate the expression quantity of transgenic plants. The results showed that the expression of transgenic tobacco with Fragment A was detected. Extracts of tobacco leaves expressing Fragment A were given to mice by gavaged, and the treated mice developed both serum IgG and secretory IgA specific for FHA.