Construction of ARRB1-EGFP fusion protein and its expression in glioma cells

Objective To construct the recombinant expression vector ARRB1-EGFP, identify its expression in cells, and lay foundations for further functional study. Methods The full gene sequence encoding human ARRB1 was obtained by RT-PCR and subcloned into the eukaryotic expression vector pEGFPN1. Recombinant expression vector, ARRB1-EGFP, was transformed into E.coli DH5α and screened by restriction enzyme digest, gel electrophoresis and DNA sequence. Then the expression vectors were transfected into HEK293 cells, and its expression products were detected by Western Blot. The subcellular localization between endogenous ARRB1 and ARRB1-GFP in the SNB19 cells was detected and compared by using immunofluorescence staining. Results ARRB1-EGFP fusion protein eukaryotic expression vector was successfully constructed, and identified by Western Blot. Localization of the fusion protein, ARRB1-GFP, was similar with that of the endogenous ARRB1. Conclusion ARRB1-EGFP fusion protein can be safely and efficiently transfected into HEK293 cells and SNB19.