AIM:To construct eukaryotic expression vector carrying human TRBP(TAR RNA-binding prorein)gene and express it in HEK-293 cells.METHODS:RT-PCR was used to obtain human full-length TRBP cDNA,Then it was cloned into the eukaryotic expression vector pFLAG-CMV4.After transfection into HEK-293 cells through lipofectamine 2000~(TM) transiently,the expression of target gene was detected by indirect immunofluorescence assay and Western Blot.RESULTS:The obtained human TRBP cDNA was consisitent with that in GenBank.The eukaryotic expression vector encoding TRBP was successfully constructed and expressed in HEK-293 cells.After eleetrophosis a protein band with molecular weight about 46 000 was found in the expression product,which is accordant with interest protein,and this band could be specifically recognized by anti-TRBP polyclonal antibody and anti-FLAG monoclona]antibody.CONCLUSION:The establishment of mammalian expression system of TRBP is of significance in further research for TRBP.
TAR RNA-binding proreinpFLAG-CMV4gene expressionHEK-293 cells
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