The Preliminary Study On Edible Vaccine Of F.hepatica Based On Transgenic Alfalfa. Containing Protective Antigenic Gene FH3

Fasciolasis is a serious parasitic disease over the world. Fosciola hepatica widely parasite in many kinds of economic herbivores such as sheep, yak, cattle, horse, deer, camel, ...etc., and hinder the development of the productive of the livestock husbandry. Moreover, the economic losses caused by fascioliasis can be very huge. So it is very important to develop an effective vaccine against fasciolisis. With the rapid development in the field of edible vaccine (oral vaccine), it becomes an available approach to use recombinant DNA methods to generate a cheap and plentiful supply of antigens. We report here the preliminary study on edible vaccine of Medicago sativa L. containing protective antigenic gene FH3 of F. hepatica.Antigenic gene FH3 of Fasciola hepatica was cloned into vector pBI121, recombinant plant expression vector was gotten, named pBI121-FH3. Agrobacteriwn tumefaciens EHA105 was transformed using pBI121-FH3. Positive EHA105 strains containing gene FH3 were picked out by plating. FH3 was introduced into alfalfa cells by co-cultivating alfalfa leaf discs with positive EHA105. Transgenic alfalfa plants were selected or gotten through methods such as tissue culture and selected culture. About 80% transgenic alfalfa plants were found containing FH3 gene by PCR and Southern blotting. Among these transgenic alfalfa plants, 5-10% plants were confirmed expressing FH3 recombinant protein, which molecular weight was about 22KD through SDS-PAGE. Based on ELISA and Western blotting, results showed that recombinant protein expressed by transgenic alfalfa plants could make responses to F. hepatica rabbit antisera.This project could explore out a new approach for studying F. hepatica plant edible vaccine.