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Expression of Colorectal Cancer Antigenic Protein Fused to IgM Fc in Chinese Cabbage (Brassica rapa).


ABSTRACT: 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 Agrobacterium-mediated transformation in Chinese cabbage (Brassica rapa). 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 Agrobacterium. The transgenic Chinese cabbages were screened using genomic polymerase chain reaction (PCR) in T0 transgenic plant lines generated from both transformants. Similarly, the immunoblot analysis revealed the expression of recombinant proteins in the transformants. Further, the T1 transgenic plants were generated by selfing the transgenic plants (T0) carrying EpCAM-IgM Fc and J-chain K proteins, respectively. Subsequently, the T1 plants generated from EpCAM-IgM Fc and J-chain K transformants were crossed to generate F1 plants carrying both transgenes. The presence of both transgenes was validated using PCR in the F1 plants. In addition, the expression of Chinese cabbage-derived EpCAM-IgM Fc × J-chain K was evaluated using immunoblot and ELISA analyses in the F1 plants. The outcomes of the present study can be utilized for the development of a potential anti-cancer vaccine candidate using Chinese cabbage.

SUBMITTER: Lee YR 

PROVIDER: S-EPMC7693566 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature

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Expression of Colorectal Cancer Antigenic Protein Fused to IgM Fc in Chinese Cabbage (<i>Brassica rapa</i>).

Lee Ye-Rin YR   Lim Chae-Yeon CY   Lim Sohee S   Park Se Ra SR   Hong Jong-Pil JP   Kim Jinhee J   Lee Hye-Eun HE   Ko Kisung K   Kim Do-Sun DS  

Plants (Basel, Switzerland) 20201030 11


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  ...[more]

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