Objective    To construct the recombinant eukaryotic expression vector pcDNA3.1(+)-β

3

-AR and express it in HEK293 cells.    Methods    Human β

3

-AR cDNA was amplified by PCR from a pENTR_ADRB3-Stop plasmid

and the eukaryotic expression plasmid pcDNA3.1(+)-β

3

-AR was constructed by inserting the β

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-AR cDNA into KpnⅠ/XbaⅠ-digested pcDNA3.1(+). The DNA sequence was confirmed

by double digestion

and the pcDNA3.1(+)-β

3

-AR plasmid was transfected into HEK293 cell line. Human β

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-AR mRNA expression on HEK293 cells was detected by RT-PCR

and the protein expression on HEK293 cells was detected by immunofluorescence technique.    Results    Restriction analysis and DNA sequencing showed that the recombinant plasmid contained the whole coding region of human β

3

-AR gene. RT-PCR assay indicated the transcription of β

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-AR mRNA and the β

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-AR protein detectable in the transfected HEK293 cells.     Conclusion    Recombinant eukaryotic expression vector pcDNA3.1(+)-β

3

-AR plasmid was successfully constructed by genetic engineering technique and expressed on the surface of HEK293 cells

which will contribute to further studies on the establishment of stable expression cell lines for screening high potent and selective β

3

-AR agonists.