[Construction of eukaryotic expression vector for HPC2 and its expression in HEK293 cells]

Di Yi Jun Yi Da Xue Xue Bao. 2005 Dec;25(12):1482-4, 1497.
[Article in Chinese]

Abstract

Objective: To construct the eukaryotic expression vector for HPC2 for expression in HEK293 cells.

Methods: HPC2 from pcDNA3/HPC2 were inserted into the flag-tagged vector pcDNA3-flag by subcloning method. The recombinant plasmid pcDNA3-flag/HPC2 was then transfected into HEK293 cells using a routine lipofectamine method. The cell lysate was used for Western blotting to examine the expression of the target protein.

Results and conclusion: Double restriction enzyme digestion and DNA sequencing indicated successful construction of the eukaryotic expression vector for HPC2 and the fusion protein was highly expressed in HEK293 cells, which provides an important basis for functional study of HPC2.

MeSH terms

  • Cell Line
  • Cloning, Molecular
  • Eukaryotic Cells / metabolism*
  • Genetic Vectors*
  • Humans
  • Ligases
  • Plasmids / genetics
  • Polycomb-Group Proteins
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics
  • Repressor Proteins / biosynthesis*
  • Repressor Proteins / genetics
  • Ubiquitin-Protein Ligases

Substances

  • Polycomb-Group Proteins
  • Recombinant Fusion Proteins
  • Repressor Proteins
  • Ubiquitin-Protein Ligases
  • Ligases
  • CBX4 protein, human