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

Title: Expression of Colorectal Cancer Antigenic Protein Fused to IgM Fc in Chinese Cabbage ( Brassica rapa )
Authors: Ye-Rin Lee; Chae-Yeon Lim; Sohee Lim; Se Ra Park; Jong-Pil Hong; Jinhee Kim; Hye-Eun Lee; Kisung Ko; Do-Sun Kim
Source: Plants, Vol 9, Iss 1466, p 1466 (2020)
Publisher Information: MDPI AG
Publication Year: 2020
Collection: Directory of Open Access Journals: DOAJ Articles
Subject Terms: Agrobacterium; colorectal carcinoma; EpCAM; plant molecular biopharming; recombinant vaccine; transgenic chinese cabbage; Botany; QK1-989
Description: 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 T 0 transgenic plant lines generated from both transformants. Similarly, the immunoblot analysis revealed the expression of recombinant proteins in the transformants. Further, the T 1 transgenic plants were generated by selfing the transgenic plants (T 0 ) carrying EpCAM–IgM Fc and J-chain K proteins, respectively. Subsequently, the T 1 plants generated from EpCAM–IgM Fc and J-chain K transformants were crossed to generate F 1 plants carrying both transgenes. The presence of both transgenes was validated using PCR in the F 1 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 1 plants. The outcomes of the present study can be utilized for the development of a potential anti-cancer vaccine candidate using Chinese cabbage.
Document Type: article in journal/newspaper
Language: English
Relation: https://www.mdpi.com/2223-7747/9/11/1466; https://doaj.org/toc/2223-7747; https://doaj.org/article/e4989fd3b11f442ca748ff47f377ec50
DOI: 10.3390/plants9111466
Availability: https://doi.org/10.3390/plants9111466; https://doaj.org/article/e4989fd3b11f442ca748ff47f377ec50
Accession Number: edsbas.EEDCB86C
Database: BASE