Eshita Yuki, Higashihara Junko, Onishi Masayasu, Mizuno Masaaki, Yoshida Jun, Takasaki Tomohiko, Kubota Naoji, Onishi Yasuhiko
Department of Infectious Disease Control, Faculty of Medicine, Oita University, 1-1 Idaigaoka, Hasama-machi, Yufu-shi, Oita Prefecture 879-5593, Japan.
Molecules. 2009 Jul 23;14(7):2669-83. doi: 10.3390/molecules14072669.
Comparative investigations were carried out regarding the efficiency of introduction of exogenous genes into cultured cells using a cationic polysaccharide DEAE-dextran-MMA (methyl methacrylate ester) graft copolymer (2-diethylaminoethyl-dextran-methyl methacrylate graft copolymer; DDMC) as a nonviral carrier for gene introduction. The results confirmed that the gene introduction efficiency was improved with DDMC relative to DEAE-dextran. Comparative investigations were carried out using various concentrations of DDMC and DNA in the introduction of DNA encoding luciferase (pGL3 control vector; Promega) into COS-7 cells derived from African green monkey kidney cells. The complex formation reaction is thought to be directly proportional to the transformation rate, but the complex formation reaction between DDMC and DNA is significantly influenced by hydrophobic bonding strength along with hydrogen bonding strength and Coulomb forces due to the hydrophobicity of the grafted MMA sections. It is thought that the reaction is a Michaelis-Menten type complex formation reaction described by the following equation: Complex amount = K1 (DNA concentration)(DDMC concentration). In support of this equation, it was confirmed that the amount of formed complex was proportional to the RLU value.
进行了比较研究，以探讨使用阳离子多糖DEAE-葡聚糖-MMA（甲基丙烯酸甲酯酯）接枝共聚物（2-二乙氨基乙基-葡聚糖-甲基丙烯酸甲酯接枝共聚物；DDMC）作为基因导入的非病毒载体，将外源基因导入培养细胞的效率。结果证实，与DEAE-葡聚糖相比，DDMC提高了基因导入效率。在将编码荧光素酶的DNA（pGL3对照载体；Promega）导入源自非洲绿猴肾细胞的COS-7细胞的过程中，使用了不同浓度的DDMC和DNA进行了比较研究。复合物形成反应被认为与转化率成正比，但由于接枝的MMA部分的疏水性，DDMC与DNA之间的复合物形成反应受到疏水键强度以及氢键强度和库仑力的显著影响。据认为，该反应是由以下方程描述的米氏型复合物形成反应：复合物量 = K1（DNA浓度）（DDMC浓度）。为支持该方程，证实形成的复合物量与相对发光单位（RLU）值成正比。
