Earticle

초록

영어

Mushroom lectins harboring carbohydrate binding specificity are powerful tools to detect glycoconjugates with N-linked or O-linked glycans for diagnosis. However, conventional preparation of diverse lectins and chemical labeling of target proteins remains a bottleneck. Here, we report a cell-free platform system for a recombinant fluorescent lectin by Escherichia coli extract in ExiProgen. Codon-optimized genes encoding the eight different types of mushroom lectins were successfully expressed as green fluorescent protein-fused forms in one-pot cell-free protein synthesis. Although the production yields of the recombinant proteins were depended on their coding genes, the resulting recombinant fluorescent lectins are apparent homo-multicomplex proteins with different molecular sizes and pI values. In hemagglutination inhibition assay, the lectins demonstrated agglutination activities towards various glycoconjugates. Moreover, GFP-fused lectins can be appliable to detect glycan binding specifies on using a glycan microarray. The glycan binding specificities of the flurescent lectins are also useful to analyze glycan epitopes expressed in different cancer cell lines. Taken together, these results provide an efficient and optimized procedure for the high-throughput synthesis and screening of lectin coding gene products based on in vitro production of the recombinant fluoresecnt lectins.

저자

• Seonghun Kim [ Jeonbuk Branch Institute, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Department of Biosystems and Bioengineering, KRIBB School of Biotechnology, University of Science and Technology (UST), 217 Gajeong-ro, Daejeon 34113, Korea ] Corresponding Author

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발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  한국당과학회 학술대회

• 간기

  연간

• 수록기간

  2006~2022

• 십진분류

  KDC 517 DDC 614