Aptamer-linked immobilized sorbent assay for detecting GMO marker, phosphinothricin acetyltransferase (PAT)

Abstract

Background

Development of genetically modified crops has rapidly increased in last few years. The most widely grown GM crops express genes that confer herbicide tolerance and insect resistance. Detection system of GM crops is important for safety evaluation before its consumption.

Objective

The purpose of this research is to detect GM crops, especially PAT, in food-samples.

Results

The bar gene (PAT protein, herbicide resistant) was cloned in pGEX-4T-1 and expressed by E. coli. The high-affinity PAT-specific single-stranded DNA (ssDNA) aptamers were obtained from a random DNA library. MOE docking study was performed to identify the potential binding region of the selected aptamers on PAT. Aptamer-linked immobilized sorbent assay (ALISA) method was used to detect PAT.

Conclusion

We screened aptamer against PAT for developing an efficient detection method. The selected PAT specific aptamers, HRPA-05 and HRPA-07, showed the distinct target binding behaviors, and detected PAT protein by aptamer-linked immobilized sorbent assay method with high efficiency and selectivity.

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Acknowledgements

This work was supported by the National Research Foundation of Korea (NRF) Grant funded by the Korea government (MEST) (no. NRF-2019R1A2C1010860).

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Contributions

Investigation and data analysis, WRS and MJL: experiments design and results analysis; JHK, SSS, JYA, YHK, SCK: Molecular docking analysis; BKC, YHK: All authors have read and agreed to the published version of the manuscript.

Corresponding authors

Correspondence to Byung-Kwan Cho or Ji-Young Ahn or Yang-Hoon Kim.

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The authors declare that they have no conflict of interest.

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Shin, W., Lee, M., Sekhon, S.S. et al. Aptamer-linked immobilized sorbent assay for detecting GMO marker, phosphinothricin acetyltransferase (PAT). Mol. Cell. Toxicol. 16, 253–261 (2020). https://doi.org/10.1007/s13273-020-00087-5

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Keywords

  • Genetically modified organism (GMO)
  • Phosphinothricin acetyltransferase (PAT)
  • ssDNA aptamer
  • Aptamer-linked immobilized sorbent assay (ALISA)