Cottrez F, Auriault C, Capron A, Kusznier J P, Groux H
Unite CNRS-URA 1854, Institut Pasteur de Lille, France.
Eur Cytokine Netw. 1994 Sep-Oct;5(5):481-7.
We described a baculovirus expression system for high level production of secreted murine recombinant IL-4. We have constructed a recombinant baculovirus based on Autographa californica polyhedrosis virus, containing both a synthetic PCR-derived murine IL-4 cDNA under the control of the polyhedrin promoter and the lacZ gene under the control of the P10 promoter to allow an easy detection of recombinant virus. The baculovirus IL-4 was fully functional in biological assay and was present under two glycosylated forms in the supernatants of infected Sf9 cells. We also detected a third unglycosylated intracytoplasmic form resulting from a fusion between the 35 first amino acids of polyhedrin and the murine IL-4. Finally, confocal microscopy showed that this recombinant protein was secreted along a classical pathway like in mammalian cells.
我们描述了一种用于高效分泌产生鼠重组白细胞介素-4(IL-4)的杆状病毒表达系统。我们构建了一种基于苜蓿银纹夜蛾核型多角体病毒的重组杆状病毒,其包含在多角体蛋白启动子控制下的合成PCR衍生的鼠IL-4 cDNA以及在P10启动子控制下的lacZ基因,以便于检测重组病毒。杆状病毒IL-4在生物学测定中具有完全功能,并且以两种糖基化形式存在于感染的Sf9细胞的上清液中。我们还检测到第三种非糖基化的胞质内形式,它是由多角体蛋白的前35个氨基酸与鼠IL-4融合产生的。最后,共聚焦显微镜显示这种重组蛋白像在哺乳动物细胞中一样沿着经典途径分泌。