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Mammalian Expression Vector Construction and the Coordinated Expression of Proteins

机译：哺乳动物表达载体构建及蛋白质的协同表达

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The goal of the project was to design and construct a recombinant expression vector that would direct the production of proteins of health interest in cultured mammalian cells. It was intended that this expression vector could direct the production of two different proteins simultaneously, in such a way that the level of production of each protein could be regulated independently. Using two existing expression vectors, the new expression vectors were constructed by rearrangement of regions of DNA using recombinant DNA technology, and tested with reporter genes, which direct the production of proteins for which the level of production in cultured mammalian cells could be measured readily. Cultured Chinese hamster ovary cells were obtained for protein production, and conditions for growth were optimized. Optimum conditions for the introduction of the expression vectors into the mammalian cells were determined. The test expression vectors containing reporting genes were introduced into mammalian cells, and production of protein was measured. Both short term transfers and permanent incorporation of the expression vectors into the cultured mammalian cells were performed, but the levels of protein production directed by the expression plasmids were found to be too low to serve as an efficient manufacturing device.

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