Objective:To construct mouse ghrelin receptor (GHS-R) eukaryotic expression vector, and observe its expression in transient transfeted COS-7 cell line.
Methods :The coding DNA sequence of ghrelin receptor gene(CDS) was amplified by SOE-PCR technology from mouse genomic DNA and cloned into PCDNA3 vector. The positive clone was confirmed by enzyme digestion and sequencing. The expression vector was transiently transfected into COS-7 cell line. The expression of GHS-R protein was identified by Western blot.
Results:The CDS of mouse GHS-R was successfully cloned. The construction of pcDNA3-GHS-R was correct and its expression of GHS-R protein was confirmed by Western blot.
Conclusions:The eukaryotic expression vector pcDNA3-GHS-R was successfully constructed and mouse GHS-R protein was successfully expressed. This study can be a basis for further study on GHS-R gene function in vitro.