Cloning of human GLIPR-2 gene and its expression in Escherichia coli

With the aim of setting a foundation for research of GLIPR2 gene function,we constructed pQE80L-GLIPR-2 vector and expressed the RGS·His-GLIPR-2 fusion protein in Escherichia coli DH5α.Firstly,the cDNA of GLIPR-2 was amplified from total RNA which was extracted from the peripheral blood by RT-PCR,and then the cDNA was inserted into pQE80L vector to construct pQE80L-GLIPR-2.Under the induction of IPTG,the GLIPR-2 and RGS·His expressed efficiently in the mode of RGS·His- GLIPR- 2 fusion protein and its expression level was analyzed by Western blotting.Cells were lysed by multiple rounds of sonication,and then the expression product was analyzed by using SDS-PAGE.The molecular weight of pQE80L- GLIPR-2 expressed in E.coli was 18 00,consistent with the expectation.The solubility analysis of expression product indicated that this recombinant protein could be expressed in both supernatant and inclusion bodies.All the result indicates that the constructed recombinant plasmid can express the soluble fusion protein efficiently in E.coli DH5α,which would be the first step for studying the GLIPR-2.