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《Journal of Tropical Medicine》 2009-04
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Cultivation and Identification of the Echinococcus granulosus Eg95-EgA31 Transgenic Alfalfa

ZHOU Hui, LI Wen-gui, YE Yan-ju, ZHOU Bi-ying (Institute of Infectious and Parasitic Diseases, the First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China)  
Objective To cultivate and identify the Echinococcus granulosus Eg95-EgA31 transgenic alfalfa. Method The recombinant pBI-Eg95-EgA31 plasmid was introduced into Agrobacterium tumefaciens by electroporation to construct recombinant Agrobacterium tumefaciens (rAt). The alfalfa leaves were infected with the recombinant Agrobacterium tumefaciens(rAt), and the somatic embryo was picked up from the callus with kanamycin resistance. Transgenic alfalfa was onstined from the resistant somatic embryo.The expression of Eg95-EgA31 fusion protein was confirmed by SDS-PAGE and Western-blot assay. The gene was identified by PCR and RT-PCR assay. Result The Eg95-EgA31 fusion gene could be expressed in alfalfa. The expression efficiency was about 0.05% of the total alfalfa leave protein. The expression product with an approximately of 37500 Mr could be recognized by sera from mice infected with E.granulosus. These were confirmed by the SDS-PAGE and Western-blot assay. The 1016bp gene was amplified successfully by PCR and RT-PCR. Conclusion The transgenic alfalfa was constructed successfully. It could provide the basis for further research of the transgenic alfalfa vaccine of Echinococcus granulosus.
【Fund】: 国家自然科学基金(No.30671835)
【CateGory Index】: S541.9
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