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[Construction of SOD1 eukaryotic expression vector and its expression].

Xiong-Hui Wu1, Hong Sun, Xiao-Wei Xing

  • 1Department of Otolaryngology, Head and Neck Surgery, Third Xiangya Hospital of Central South University, Changsha 410013, China. wuxh-2008@163.com

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi = Chinese Journal of Cellular and Molecular Immunology
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Summary

A new eukaryotic expression vector for human superoxide dismutase 1 (SOD1) was successfully constructed. This vector enables the expression of SOD1 and green fluorescent protein (GFP) in HeLa cells, offering a tool for gene therapy research.

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Area of Science:

  • Molecular Biology
  • Gene Expression
  • Cell Biology

Context:

  • Human Cu/Zn superoxide dismutase 1 (SOD1) plays a critical role in cellular defense against oxidative stress.
  • Developing efficient tools for studying SOD1 expression is crucial for understanding its function and potential therapeutic applications.

Purpose:

  • To construct a novel eukaryotic expression vector for human SOD1.
  • To validate the expression of SOD1 and a reporter gene (GFP) in HeLa cells using the constructed vector.

Summary:

  • The open reading frame of human SOD1 was amplified and cloned into the pUCm-T vector, creating pUCm-T-SOD1.
  • SOD1 was subsequently subcloned into the eukaryotic expression vector pTracer-CMV/Bsd, yielding pTracer-CMV/Bsd-SOD1.
  • Transfection into HeLa cells via Lipofectamine 2000 resulted in observable GFP expression and detectable SOD1 expression via RT-PCR and Western blot after blasticidin selection.

Impact:

  • Successful construction of the pTracer-CMV/Bsd-SOD1 vector provides a valuable tool for gene therapy studies.
  • This vector facilitates the simultaneous expression of SOD1 and GFP, enabling robust monitoring of gene delivery and expression in eukaryotic cells.
  • The findings support further investigation into SOD1's role in disease and its therapeutic potential.