Abstract:Objective:To construct a eukaryotic expressing vector harboring human melanin-concentrating hormone receptor 2 (MCHR2) and to establish a SHG-44 cell line stably and highly expressing MCHR2.Methods:The full-length MCHR2 cDNA fragment was amplified from the human fetal brain cDNA library by PCR and was cloned into pcDNA3.1(+) to construct eukaryotic vector pcDNA3.1(+)/MCHR2; the latter was then transduced into SHG-44 cells by LipofectamineTM. After screening culture by G418,SHG-44 cells stably expressing MCHR2 were established. The transcription and expression of MCHR2 was identified by RT-PCR,Western blotting and immunofluorescence.Results:The full-length MCHR2 cDNA fragment was amplified and the eukaryotic expression vector pcDNA3.1 (+)/MCHR2 was successfully constructed.The expression of MCHR2 was found positive by RT-PCR,Western blotting and immunofluorescence,indicating that the SHG-44 cell line stably and highly expressing MCHR2 was successfully established.Conclusion:The successful establishment of MCHR2-SHG-44 cell line provides a solid foundation for further study on MCHR2 function.