Construction of eukaryotic expression vector of human MCHR2 and establishment of its stable transfected CHO cell line

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.