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Cloning and Expression of Oil Palm (Elaeis guineensis Jacq.) Type 2 Ribosome Inactivating Protein in Escherichia coli

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Abstract

EgT2RIP is a type 2 ribosome-inactivating protein isolated from oil palm (Elaeis guineensis Jacq.). Its transcript abundance was reported to be up-regulated in oil palm roots upon inoculation of pathogenic fungus Ganoderma boninense in a recent study. This study aims to produce an active recombinant EgT2RIP protein for biological studies. The DNA fragments encoding Chain A (CA) and Chain B (CB) of EgT2RIP were cloned individually in an expression vector. Soluble CA and partially soluble CB were expressed in Escherichia coli Rosetta-gami 2 (DE3). Purified recombinant CA and CB were associated in a cysteine/cystine reduced/oxidized system, yielding a heterodimer protein (AB). The AB protein showed growth inhibitory activity against breast cancer cell lines (MCF-7) as well as non-tumorigenic breast epithelial cell line (MCF-10A) at IC50 = 1.4 and 10.9 μg mL−1, respectively. The active protein produced from this study may have the potential to be used for treatment in medical and agricultural fields.

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Abbreviations

AB:

Heterodimers of Chains A and B

CA:

Chain A

CB:

Chain B

EST:

Expressed sequence tag

RIP:

Ribosome inactivating protein

SDS–PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

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Acknowledgments

This project is funded by RUGS Initiative 6 (Grant No: 05-02-11-1408RU). We thank the Pathology Laboratory of Malaysian Palm Oil Board (MPOB, Malaysia) for providing the G. boninense PER71. Yung-Chie Tan was supported by Ministry of Science, Technology and Innovation (MOSTI) Malaysia under the National Science Fellowship (NSF).

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Authors and Affiliations

  1. Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, UPM, 43400, Serdang, Selangor, Malaysia

    Noorjahan Banu Alitheen & Chai-Ling Ho

  2. Institute of Tropical Agriculture, Universiti Putra Malaysia, UPM, 43400, Serdang, Selangor, Malaysia

    Yung-Chie Tan, Mui-Yun Wong & Chai-Ling Ho

  3. Department of Biomedical Sciences, University of Nottingham Malaysia Campus, Semenyih, Selangor, Malaysia

    Wan-Yong Ho

  4. Department of Plant Protection, Faculty of Agriculture, Universiti Putra Malaysia, UPM, 43400, Serdang, Selangor, Malaysia

    Mui-Yun Wong

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  1. Yung-Chie Tan
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  2. Wan-Yong Ho
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  3. Noorjahan Banu Alitheen
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  4. Mui-Yun Wong
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  5. Chai-Ling Ho
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Correspondence to Chai-Ling Ho.

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This article does not contain any studies with human or animal subjects performed by any of the authors.

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10989_2015_9485_MOESM1_ESM.docx

Supplementary material 1. Supplementary Fig. A Nucleotide sequence and deduced amino acid sequence of EgT2RIP. The predicted N-terminal signal peptide and linker peptide are underlined. The position of the putative translation termination codon is indicated by an asterisk (DOCX 17 kb)

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Tan, YC., Ho, WY., Alitheen, N.B. et al. Cloning and Expression of Oil Palm (Elaeis guineensis Jacq.) Type 2 Ribosome Inactivating Protein in Escherichia coli . Int J Pept Res Ther 22, 37–44 (2016). https://doi.org/10.1007/s10989-015-9485-5

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  • DOI: https://doi.org/10.1007/s10989-015-9485-5

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