Polycation graft copolymers accelerating DNA strand exchange: involvement of ionic interaction.
In the previous study (Chem. Eur. J., 7, 176 (2001)) we demonstrated that the comb-type polycationic copolymer (PLL-g-Dex) which is composed of poly (L-lysine) backbone and dextran graft chains expedited the DNA strand exchange reaction. In this study, fluorescence resonance energy transfer (FRET) was employed to explore the copolymer-mediated DNA strand exchange with higher time-resolution. To initiate strand exchange reaction the duplex prepared from 3'-fluorescein isothiocyanate (FITC)--and 5'-carboxytetramethylrhodamine(TAMRA)-labeled complementary DNAs was added to its non-labeled complementary single strand. DNA strand exchange was monitored by observing the recovery of the FITC quenching. More than 20,000 times increase in strand exchange rate at 37 degrees C by the copolymer was estimated. To investigate the accelerating mechanisms of the copolymer, the same reactions but at various ionic strenghts were studied. With increasing ionic strengths the strand exchange rate in the absence of the copolymer increased, suggesting that ionic repulsion among DNAs is unfavorable for the strand exchange to occur. Hence, alleviation of the electrostatic repulsion through interpolyelectrolyte complex formation is probably a role of the copolymer for accelerating the strand exchange reaction.
Kim
W J
WJ
Department of Biomolecular Engineering, Tokyo Institute of Technology, 4259 Nagatsuta, Midori, Yokohama 226-8501, Japan.
Ishihara
T
T
Akaike
T
T
Maruyama
A
A
D004247:DNA / Q000737:chemistry / Q000187:drug effects* / Q000235:genetics; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000235:genetics; D003911:Dextrans / Q000737:chemistry / Q000494:pharmacology*; D031541:Fluorescence Resonance Energy Transfer; D007477:Ions; D007700:Kinetics; D011073:Polyamines / Q000737:chemistry / Q000494:pharmacology; D000071228:Polyelectrolytes; D011107:Polylysine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology*; D011995:Recombination, Genetic / Q000187:drug effects*; D012965:Sodium Chloride / Q000494:pharmacology
D004247:DNA; D004277:DNA, Single-Stranded; D003911:Dextrans; D007477:Ions; D011073:Polyamines; D000071228:Polyelectrolytes; D011107:Polylysine; D012965:Sodium Chloride; C113090:poly(lysine)-graft-dextran