家蚕中一个新的膜蛋白基因HW的融合表达与纯化

HW gene was found from the cDNA library constructed by our laboratory. It is 1230 bp long and has an open reading frame of 294 bp, encoding a protein of 97 amino acid residues with a predicted molecular weight of 10.93 kD. HW gene was fused with Bombyx mori baculovirus polyhedrin gene. This fused gene was inserted into the vector pET-28a(+) to construct the recombinant expression plasmid pET-28a-Ph-HW and expressed in E.coli. We purified the fused protein by dissolving inclusion ptotein in different pH buffer solution and metal-chelating affinity chromatography. At the same time, the fused gene was inserted into BmBacmid and transfected into BmN cells at the base of Bac-to-Bac system. The recombinant BmBacmid-Ph-HW was obtained and expressed successfully. This new silkworm membrane protein HW was expressed in E.coli and silkworm cell successfully and the expression level was high in E.coli.The fused polyhedrin protein promoted the expresson of membrane protein HW and made the purification of fusion protein easy. This should lay a good basis for researching the structure and function of this transmenbrane protein HW.