Kameda S, Maruyama H, Higuchi N, Nakamura G, Iino N, Nishikawa Y, Miyazaki J, Gejyo F
Division of Clinical Nephrology and Rheumatology, Niigata University Graduate School of Medicine and Dental Sciences, 1-757 Asahimachi-dori, Niigata 951-8120, Japan.
Biochem Biophys Res Commun. 2003 Oct 3;309(4):929-36. doi: 10.1016/j.bbrc.2003.08.087.
A high level of plasmid DNA expression in rat liver can be achieved by the rapid injection of a large volume of a naked DNA solution into the tail vein, called the 'hydrodynamics-based procedure.' The preparation of PCR-amplified DNA fragments is easier than that of naked DNA. In this paper we evaluated the effects of expressing the erythropoietin (Epo) gene in the rat liver by injecting fCAGGS-Epo, an Epo-expressing PCR-amplified DNA fragment, via the tail vein. After injection of 5 pmol fCAGGS-Epo (10 microg) or pCAGGS-Epo (18.4 microg), plasmid DNA, the serum Epo levels peaked at week 1, then persisted for at least 12 weeks. Transgene-derived Epo secretion resulted in significant erythropoiesis. These results demonstrated that transfer of PCR-amplified DNA fragments into the rat liver via rapid tail vein injection can be achieved. This method may provide a useful means for studying the physiologic function of a putative gene.
通过向大鼠尾静脉快速注射大量裸DNA溶液（即“基于流体动力学的方法”），可在大鼠肝脏中实现高水平的质粒DNA表达。PCR扩增DNA片段的制备比裸DNA更容易。在本文中，我们评估了通过尾静脉注射fCAGGS-Epo（一种表达促红细胞生成素的PCR扩增DNA片段）在大鼠肝脏中表达促红细胞生成素（Epo）基因的效果。注射5 pmol fCAGGS-Epo（10微克）或pCAGGS-Epo（18.4微克）质粒DNA后，血清Epo水平在第1周达到峰值，然后持续至少12周。转基因衍生的Epo分泌导致显著的红细胞生成。这些结果表明，通过快速尾静脉注射将PCR扩增的DNA片段转移到大鼠肝脏中是可行的。该方法可能为研究假定基因的生理功能提供一种有用的手段。
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