Establishment of CHO cell line stably expressing cannabinoid receptor 2

AIM To establish Chinese hamster ovary(CHO) cell strains stably expressing human cannabinoid receptor 2(CB2).METHODS Plasmid vector pcDNA3.1(+)-CB2 that contains a cDNA fragment coding for human CB2 was transfected into CHO cells by a Lipofectamine based method,and transfected CHO cells were selected in culture medium containing G418.Positive cell clones were measured by RT-PCR.The DNA fragment was sequenced.The activity of CB2 was analyzed by radioligand receptor binding assays.RESULTS Through screening by G418,twenty-eight CHO cell monoclones were obtained.Seventeen positive CHO cell clones expressed CB2 mRNA analyzed by RT-PCR,and the sequence of the PCR fragment of CB2 was right.The saturation analysis indicated that the CB2 receptor of the recombinants CHO cell strain should be similar to the natural CB2.The value of Kd and Bmax was(1.21±0.47)nmol·L-1 and(3.12±0.7)nmol·g-1 protein,respectively.CONCLUSION CHO cell stains stably expressing human CB2 with activity are established.