Constructing the recombinant of the humanbeings' GOR gene and its expression within Cos-7 cell line

Objective To construct eukariotic recombinants able to express human beings' GOR protein and to abtain the protein. Methods The fragments of human being's GOR gene were inserted to the vector system of pcDNA3.1(-)/Myc-His A, B and C, and then positive recombinants were selected by the technique of PCR and EcoR I digestion. The recombinants with sense sequense were determined by Kpn I digestion. After three kinds of recombinant transfecting Cos-7 cells with the calcium phosphate precipitation method respectively, the technique of PCR and ELISA were used to confirm whether the recombinants could be transcripted and expressed or not within cells. Plenty of Cos-7 cells cultured and transfected with recombinant-A, the protein of human beings' GOR was got. Results All of the three kinds of recombinant A, B, C could be transcripted within Cos-7 cells, but only recombinant A could express human beings'GOR protein. Conclusion Eukariotic recombinants constructed could exactly express human beings' GOR protein. [