In vitro transfection of plasmid DNA by cationized gelatin prepared from different amine compounds
In vitro transfection of plasmid DNA by cationized gelatin prepared from different amine compounds
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DOI：
10.1163/156856206777346278
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发表时间：
2006-01-01
影响因子：
3.6
通讯作者：
Tabata, Yasuhiko
中科院分区：
文献类型：
--
作者：
Kushibiki, Toshihiro;Tomoshige, Ryuji;Tabata, Yasuhiko
比较了不同胺类化合物制备的阳离子明胶对荧光素酶质粒DNA的体外转染效率。这里使用的化合物是乙二胺，腐胺，亚精胺和精胺，化学引入明胶的羧基进行阳离子化。阳离子化明胶与质粒DNA的络合通过在水溶液中以各种N+/P-混合比（明胶的氨基与DNA的磷酸基团的摩尔数比）简单地混合两种材料来进行。凝胶阻滞研究表明，阳离子化明胶质粒DNA复合物的形成依赖于N+/P-混合比。溴化乙锭插层实验和Scatchard结合分析表明，质粒DNA与精胺阳离子明胶的相互作用较强。当对小鼠L929成纤维细胞评价以不同N+/P-混合比与各种阳离子化明胶复合的质粒DNA的转染效率时，观察到由精胺的阳离子化明胶以N+/P-混合比为2制备的复合物的最高转染效率。本研究表明，存在一个最佳的N+/P-混合比和一种胺类化合物或阳离子化明胶的阳离子化程度，以提高质粒DNA的转染效率。
The objective of this paper is to compare the in vitro transfection efficiency of a luciferase plasmid DNA using cationized gelatin prepared from different amine compounds. The compounds used here were ethylenediamine, putrescine, spermidine and spermine, chemically introduced to the carboxyl group of gelatin for the cationization. Complexation of the cationized gelatin with the plasmid DNA was performed by simply mixing the two materials at various N+/P- mixing ratios (the molar number ratio of amino groups of gelatin to the phosphate groups of DNA) in aqueous solution. Gel retardation studies revealed that the formation of cationized-gelatin-plasmid DNA complexes depended on the N+/P- mixing ratio. The stronger interaction of plasmid DNA with the cationized gelatin of spermine compared to the other cationized gelatins was observed by an ethidium bromide intercalation assay and Scatchard binding analysis. When the transfection efficiency of plasmid DNA complexed with the various cationized gelatins at different N+/P- mixing ratios was evaluated for mouse L929 fibroblasts, the highest transfection efficiency was observed for the complex prepared from the cationized gelatin of spermine at a N+/P- mixing ratio of 2. The present study indicates that there is an optimal N+/P- mixing ratio and a type of amine compound or cationization extent of cationized gelatin to enhance the transfection efficiency of plasmid DNA.