Objective     To construct eukaryotic expression vector of human melanin-concentrating hormone receptor 2 (MCHR2) and transfect CHO cells to establish stable CHO cell line.       Methods     The full-length MCHR2 cDNA fragment was amplified by PCR from the human fetal brain cDNA library and was inserted into eukaryotic expression vector pcDNA3.1(+)

after the identification by digestion and sequencing on the recombinant eukaryotic expression vector pcDNA3.1(+)/MCHR2

the recombinant was transfected into CHO cell by lipofectamine

TM

2000. After screening culture by G418

stable transfected CHO cell line was established

and the transcription and expression of MCHR2 were identified by RT-PCR

Western

blotting and immunofluorescence.      Results     The eukaryotic expression vector pcDNA3.1(+)/MCHR2 was constructed successfully. The stable transfected CHO cell line was established. The MCHR2 protein was expressed successfully.      Conclusion     The construction of the eukaryotic expression vector pcDNA3.1(+)/MCHR2 and the establishment of stable transfected CHO cell line provide solid foundation for further experimental studies on the function of MCHR2.