Abstract: The objective of this research is to construct a site-specific integration vector harboring the E2 gene of Hog Cholera Virus, the gene coding for the main antigen, for transformation of chloroplast genome of Lotus corniculatus, which is the groundwork for producing edible plant vaccines in the chloroplast of this forage. The optimal site was selected for integration of exogenous genes into the chloroplast genome of Lotus corniculatus, and primers were designed and used to amplify chloroplast DNA fragments. After cloning of these fragments, the plastid transformation vector of Lotus corniculatus was constructed. The constructed vector, the pAKE2, contains two neighboring DNA fragments of 1.35 kb and 1.5 kb in the region of psbA and trnK genes, respectively, as targeting sequences for the chloroplast transformation of Lotus corniculatus, as well as the E2 gene cassette, which is inserted between the strong chloroplast promoter Prrn and the terminator of psbA gene form tobacco, and the aadA cassette, which is used for spectinomycine resistance selection of transformants. The results of PCR amplification and restriction enzyme digestion analysis showed that the constructed vector is in full accordance with what was desired. The transformation and following works are in progress.

Key words: E2 gene of Hog Cholera Virus, chloroplast transformation, homologous recombination, site-specific integration, Lotus corniculatus