[Construction and functional study of truncated beta catenin genes].
OBJECTIVE
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.


CONCLUSIONS
The different locations of truncated beta catenins in transfected COS7 cells suggest that the truncated ones play different roles in the transcriptional activity.
Zhang
Rui
R
Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China.
Yan
He-xin
HX
Chen
Lei
L
Tang
Liang
L
Yang
An-gang
AG
Wang
Hong-yang
HY
D000818:Animals; D019556:COS Cells / Q000378:metabolism; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D004622:Embryo, Mammalian; D017353:Gene Deletion; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D014158:Transcription, Genetic; D014162:Transfection; D051176:beta Catenin
C495270:CTNNB1 protein, human; D003598:Cytoskeletal Proteins; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D051176:beta Catenin; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins