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[Construction and expression of vector encoding Sox2 with mutated SUMO accepter site].

Ze-kun Guo1, Xian-qiang Wu, Li-ran Shan

  • 1College of Life Sciences, Northwest A & F University, Yangling 712100, China. gzkster@gmail.com

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi = Chinese Journal of Cellular and Molecular Immunology
|November 11, 2009
PubMed
Summary

We successfully created and expressed both wild-type Sox2 and a mutant form (Sox2 K247R). The mutant Sox2 protein is not subject to SUMOylation, indicating a key role for lysine 247 in this post-translational modification.

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Area of Science:

  • Molecular Biology
  • Protein Biochemistry
  • Gene Expression

Context:

  • Sox2 is a crucial transcription factor involved in maintaining pluripotency and driving cell differentiation.
  • Post-translational modifications, such as SUMOylation, play significant roles in regulating protein function and stability.
  • Understanding Sox2 regulation is vital for stem cell research and regenerative medicine.

Purpose:

  • To construct eukaryotic expression plasmids for wild-type Sox2 and a specific mutant, Sox2 K247R.
  • To analyze the expression levels of both Sox2 variants in mammalian cells.
  • To investigate the SUMOylation status of Sox2 and the impact of the K247R mutation on this modification.

Summary:

  • Eukaryotic expression vectors for Sox2 and a mutant form (Sox2 K247R) were successfully constructed and verified by sequencing.
  • Western blot analysis confirmed robust expression of both wild-type and mutant Sox2 in 293FT cells.
  • The study demonstrated that wild-type Sox2 undergoes SUMOylation, while the Sox2 K247R mutant is resistant to SUMOylation, highlighting the importance of lysine 247.

Impact:

  • Provides functional plasmids for further research on Sox2.
  • Elucidates the role of lysine 247 in Sox2 SUMOylation.
  • Contributes to understanding the regulatory mechanisms of Sox2 in biological processes.