[Construction and functional study of truncated beta catenin genes]

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2004 Jul;20(4):385-9.
[Article in Chinese]

Abstract

Aim: To observe the subcellular location and the change of the transcriptional activity of truncated beta catenins in mammalian cells.

Methods: The full length wild type beta catenin gene was cloned by RT-PCR. The truncated beta catenins were constructed by deleting the N terminal, the C terminal or the Armadillo domain sequences, and were inserted into the EGFP expression vector. After they were transfected into 293 cells, the transcriptional activity of truncated beta catenins in 293 cells was detected by dual luciferase reporter system. The subcellular location of truncated beta catenin in transfected COS7 cells was observed under fluorescence microscope.

Results: The eukaryotic expression vector containing truncated beta catenin gene and expression vector of truncated beta catenin gene fused with EGFP gene were constructed successfully. C terminus-deleted beta catenin localized mainly in the cytoplasm of transfected COS7 cells, and the transcriptional activity of the N terminus- and the C terminus- deleted beta catenins was lower than that of the wild type beta catenin in transfected 293 cells.

Conclusion: The different locations of truncated beta catenins in transfected COS7 cells suggest that the truncated ones play different roles in the transcriptional activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • COS Cells / metabolism
  • Cell Line
  • Cell Nucleus / metabolism
  • Chlorocebus aethiops
  • Cloning, Molecular
  • Cytoplasm / metabolism
  • Cytoskeletal Proteins / genetics*
  • Cytoskeletal Proteins / metabolism
  • Cytoskeletal Proteins / physiology
  • Embryo, Mammalian
  • Gene Deletion*
  • Genetic Vectors
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Kidney / cytology
  • Kidney / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Trans-Activators / genetics*
  • Trans-Activators / metabolism
  • Trans-Activators / physiology
  • Transcription, Genetic*
  • Transfection
  • beta Catenin

Substances

  • CTNNB1 protein, human
  • Cytoskeletal Proteins
  • Recombinant Fusion Proteins
  • Trans-Activators
  • beta Catenin
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins