Stable expression of pcDNA3.1-hTSHR in CHO cells

Objective To construct the recombinant eukaryotic expression vector with full length cDNA sequence of pcDNA3.1/human thyroid-stimulating hormone receptor(hTSHR)(accomplished) and to express it in CHO cells(Chinese hamster ovary cells),then screen the stable expression cell lines.Methods Constructed pcDNA3.1-hTSHR was transferred into CHO cells by liposome.The stable expression cell lines were screened by using G 418 and were cultured for generations.The fifth generation was selected and DNA and protein expressions of hTSHR in transferred CHO cells were detected by PCR and Western blot.Results Five monoclones with full length sequence of hTSHR gene were obtained.One band of approximately 530 bp in size was amplified by PCR which agreed with anticipation.Western blot confirmed that the expressed recombinant hTSHR protein was present and active in CHO cells.Conclusion The stable hTSHR expression cell lines has been successfully constructed,which would contribute to further studies on the TSHR function in the blood of patients with autoimmune thyroid disease.