Related Experiment Videos
[Construction and functional study of truncated beta catenin genes]
Rui Zhang1, He-xin Yan, Lei Chen
1Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China.
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.