人M-CSF与SCF融合蛋白的构建及其在昆虫细胞中的表达

Construction and expression of human macrophage colony stimulating factor and stem cell factor fusion protein in Sf9 cells

应用重组DNA技术构建M CSF与SCF的融合基因并将其克隆于昆虫杆状病毒转移载体 pVL13 92中 ,通过与野生型苜蓿夜蛾核型多角体病毒 (AcNPV)DNA共转染草地夜蛾细胞Sf9,融合基因插入AcNPV基因组。重组病毒感染单层Sf9细胞后 ,表达产物分泌到胞外培养液中 ,用MTT比色法和TF 1细胞株可检测到表达产物与IL 3的协同效应。

A fusion gene was constructed in a plasmid in which the coding regio ns of human Macrophage Colony Stimulating Factor (M-CSF) and Stem Cell Factor ( S CF) cDNAs were connected through DNA recombinant techniques. It was then subclon ed into the baculovirus transfer vector pVL1392 with the control of polyhedrin p romoter. The Sf9 cells (Spodoptera frugiperda) were cotransfected with the recom binant plasmid and AcNPV DNA (Autographa californica nuclear polyhedrosis virus DNA) to produce the recombinant virus by homologous recombination. The Sf9 infec ted with the recombinant baculovirus expressed fusion protein which was secreted into the cell culture. The synergistic activities of SCF in conjunction with hu man interleukin-3(hIL-3) was measured by MTT colorimetric method and TF-1 cel l line.