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KMID : 0380020040190050341
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Korean Journal of Biotechnology and Bioengineering
2004 Volume.19 No. 5 p.341 ~ p.347
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Studies on Developing Direct Gene Transfer Based on Naked Plasmid DNA for Treating Anemia
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Park Young-Sup
Jung Dong-Gun
Choi Cha-Yong
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Abstract
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Several gene delivery therapies are being developed for treatment of serum protein deficiency. EPO is one of the most promising therapeutic agent for this treatment which is currently being investigated in depth. This study has the ultimate purpose of improving the gene delivery system for an increase of red blood cell production. A plasmid DNA was constructed smaller than other plasmids for an increase in penetration into animal cells, and two genes were cloned into each vector as a co-delivery system to express erythropoietin, and interluekin-3 or thrombopoietin, which can act on erythroid cell, thus activating hematopoiesis synergically. This co-delivery system has an advantage of decreasing the labour required for industrial production of DNA vaccine. A new plasmid vector, pVAC, in size 2.9 kb, was constructed with the essential parts from PUC 19 and pSectagB, which is much smaller than other plasmid vector and is the size of 2.9 kb. Co-delivery system was constituted by cloning human erythropoietin with each of human interluekin-3 gene or human thrombopoietin gene into both pVAC and pSectagB. As a result, the transfection efficiency of pVAC was higer than that of pSectagB in vitro, and hematocrit level of the mice injected with pVAC is higher than that of other mice. And co-delivery system, made of several plasmid DNAs, was expressed in vitro.
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KEYWORD
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DNA vaccine, plasmid DNA, human erythropoietin, hEPO, human interleukin3, hIL-3, human trombopoietin, hTPO, in vitro transfection, in vivo, hematocrit
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