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Abstract <p>The functioning of proteins in a cell cannot be studied without analyzing their content in the cell. The most commonly used method of analysis, Western blotting, cannot always be used due to the impossibility of obtaining antibodies that specifically recognize the protein of interest. At the same time, Western blotting is a semi-quantitative method of analysis and does not allow determining the exact content of protein in the cell. In this work, we used the protein complementation method for the qualitative and quantitative assessment of the hTERP protein content in the cell. Using genome editing, a HiBiT epitope was introduced to the C-terminus of the hTERP protein, the complementation of which with LgBiT restores active luciferase. As a result, we determined the hTERP protein content in the HEK293T cell line.</p>

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Protein Complementation Approach for Quantitative and Qualitative Analysis of hTERP in Cell

  • 期刊论文

  • 发表日期：

    2025年8月1日

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  • M. A. Shamonova,
  • M. S. Koriagina,
  • V. L. Shliapina,
  • O. A. Dontsova,
  • M. P. Rubtsova

摘要

Abstract

The functioning of proteins in a cell cannot be studied without analyzing their content in the cell. The most commonly used method of analysis, Western blotting, cannot always be used due to the impossibility of obtaining antibodies that specifically recognize the protein of interest. At the same time, Western blotting is a semi-quantitative method of analysis and does not allow determining the exact content of protein in the cell. In this work, we used the protein complementation method for the qualitative and quantitative assessment of the hTERP protein content in the cell. Using genome editing, a HiBiT epitope was introduced to the C-terminus of the hTERP protein, the complementation of which with LgBiT restores active luciferase. As a result, we determined the hTERP protein content in the HEK293T cell line.