| Summary: | The epithelial cell adhesion molecule (EpCAM) is a tumor-associated antigen and a potential target for tumor vaccine. The EpCAM is a cell-surface glycoprotein highly expressed in colorectal carcinomas. The objective of the present study is to develop an edible vaccine system through <i>Agrobacterium</i>-mediated transformation in Chinese cabbage (<i>Brassica rapa</i>). For the transformation, two plant expression vectors containing genes encoding for the EpCAM recombinant protein along with the fragment crystallizable (Fc) region of immunoglobulin M (IgM) and Joining (J)-chain tagged with the KDEL endoplasmic reticulum retention motif (J-chain K) were constructed. The vectors were successfully transformed and expressed in the Chinese cabbage individually using <i>Agrobacterium</i>. The transgenic Chinese cabbages were screened using genomic polymerase chain reaction (PCR) in T<sub>0</sub> transgenic plant lines generated from both transformants. Similarly, the immunoblot analysis revealed the expression of recombinant proteins in the transformants. Further, the T<sub>1</sub> transgenic plants were generated by selfing the transgenic plants (T<sub>0</sub>) carrying EpCAM–IgM Fc and J-chain K proteins, respectively. Subsequently, the T<sub>1</sub> plants generated from EpCAM–IgM Fc and J-chain K transformants were crossed to generate F<sub>1</sub> plants carrying both transgenes. The presence of both transgenes was validated using PCR in the F<sub>1</sub> plants. In addition, the expression of Chinese cabbage-derived EpCAM–IgM Fc × J-chain K was evaluated using immunoblot and ELISA analyses in the F<sub>1</sub> plants. The outcomes of the present study can be utilized for the development of a potential anti-cancer vaccine candidate using Chinese cabbage.
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