Secretory production of a functional membrane protein

点击翻译按钮获取中文摘要

The DNA fragment encoding the bacteriorhodopsin gene from Halobacterium halobium was cloned into a plasmid vector that contained the Bombyx mori actin promoter downstream of the B. mori nucleopolyhedrovirus (BmNPV) IE-1 transactivator and the BmNPV HR3 enhancer for high-level expression. After transfection with the resultant plasmid into Trichoplusia ni BTI-TN-5B1-4 (High Five) cells, cellular extract was analyzed by western blot analysis. Specific protein bands were detected at electrophoretic mobility of approximately 20 kDa in the cell lysate, indicating successful expression of bacteriorhodopsin in insect cells. The plasmid vector was then transfected into recombinant High Five cells that expressed the precursor (prM) of the viral membrane protein (M) and the envelope glycoprotein (E) of Japanese encephalitis (JE) virus. Secretory expression of virus-like particles embedding bacteriorhodopsin has been examined.