Cloning and Expression of the Human SH2B1β

Objective To obtain the sequence of the human SH2B1β gene, construct a recombinant expression plasmid and identify the expressed product. Methods The SH2B1β cDNA was amplified from the RNA of human differentiated fat cells by RT-PCR. The gene fragment was cloned into vector pET28a(+), then expressed in E. coli BL21(DE3). Result The result of and restriction analysis showed that SH2B1β had been cloned into pET28a (+ )vector, and the result of DNA sequence analysis also showed the cloned SH2B1β gene sequence was the same as the published sequence in GenBank database. The recombinant protein, about 75 kDa, was detected by SDS-PAGE. Conclusion The prokaryotic expression plasmid SH2B1β- pET28a (+ ) was successfully constructed and SH2B1β protein was expressed in E. coli BL21 (DE3). Our work may provide foundation for production and functional study of this protein.