Construction of thioredoxin-DsRed and APE/ref-1-EGFP fusion protein vectors and their localization in 293T cells
- VernacularTitle:硫氧还蛋白和氧化/还原因子的融合荧光蛋白载体的构建及在293T细胞中的分布
- Author:
Zhenhua XIE
- Publication Type:Journal Article
- Keywords:
thioredoxin;
APE/ref-1;
Green fluorescent protein:Red fluorescent proteins
- From:
Basic & Clinical Medicine
2006;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct thioredoxin and APE/ref-1 fusion protein expression vector and to investigate their subcellular localization of the fusion proteins in 293Tcells.Methods The APE/ref-1 cDNA was cloned by RT-PCR from PC12 cell.APE/ref-1-EGFP fusion protein expression vector was constructed through subcloning.Thioredoxin cDNA was subcloned into pDSred1-1 from pQE30-TRX plasmid by PCR,and thioredoxin-DsRed fusion protein expression vector was constructed in pCMV5 plasmid.The expression and subcellular localization of the fusion proteins in 293T cells transfected with the vectors by calcium phosphate was analyzed by fluorescent microscopy.Results The results demonstrated that thioredoxin——DsRed and APE/ref-1-EGFP fusion protein expression vectors were successfully constructed and expressed in 293T cells.APE/ref-1-EGFP fusion protein was located only in nucleus,and thioredoxin-DsRed fusion protein was located in cytoplasm as well as nucleus.ConclusionThis study has set up a solid base for further to study on the dynamic interaction between thioredoxin and APE/ref-1 fusion proteins.
