甲酰胺和尿素水溶液中DNA的复性动力学及热稳定性
Renaturation kinetics and thermal stability of DNA in aqueous solutions of formamide and urea.
作者信息
Hutton J R
出版信息
Nucleic Acids Res. 1977 Oct;4(10):3537-55. doi: 10.1093/nar/4.10.3537.
Abstract
This paper reports the results of a systematic study of the effects of formamide and urea on the thermal stability and renaturation kinetics of DNA. Increasing concentrations of urea in the range 0 to 8 molar lower the Tm by 2.25 degrees C per molar, and decreases the renaturation rate by approximately 8 percent per molar. Increasing concentrations of formamide in the range from 0 to 50 percent lowers the Tm by 0.60 degrees C per percent formamide for sodium chloride concentrations ranging from 0.035M to 0.88M. At higher salt concentrations the dependence of Tm on percent formamide was found to be slightly greater. Increasing formamide concentration decreases the renaturation rate linearly by 1.1% per percent formamide such that the optimal rate in 50% formamide is 0.45 the optimal rate in an identical solution with no formamide. The effects of urea and formamide on the renaturation rates of DNA are explained by consideration of the viscosities of the solutions at the renaturation temperatures.
摘要
本文报道了关于甲酰胺和尿素对DNA热稳定性及复性动力学影响的系统研究结果。在0至8摩尔范围内，尿素浓度的增加使熔解温度（Tm）每摩尔降低2.25摄氏度，且使复性速率每摩尔降低约8%。在氯化钠浓度为0.035M至0.88M时，甲酰胺浓度在0至50%范围内增加，使Tm每百分比甲酰胺降低0.60摄氏度。在更高盐浓度下，发现Tm对甲酰胺百分比的依赖性略大。甲酰胺浓度增加使复性速率以每百分比甲酰胺1.1%的比例线性降低，因此在50%甲酰胺中的最佳复性速率是在无甲酰胺的相同溶液中的最佳复性速率的0.45倍。通过考虑复性温度下溶液的粘度来解释尿素和甲酰胺对DNA复性速率的影响。
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本文引用的文献
1
A fractionating column for analysis of nucleic acids.用于核酸分析的分馏柱。
Anal Biochem. 1960 Jun;1:66-77. doi: 10.1016/0003-2697(60)90020-8.
2
Electrostatic effects on polynucleotide transitions. II. Behavior of titrated systems.静电对多核苷酸转变的影响。II. 滴定体系的行为
Biopolymers. 1967;5(10):993-1008. doi: 10.1002/bip.1967.360051011.
3
Electrostatic effects on polynucleotide transitions. I. Behavior at neutral pH.静电作用对多核苷酸转变的影响。I. 中性pH条件下的行为
Biopolymers. 1967;5(10):975-92. doi: 10.1002/bip.1967.360051010.
4
Dependence of the melting temperature of DNA on salt concentration.DNA熔解温度对盐浓度的依赖性。
Biopolymers. 1965;3(2):195-208. doi: 10.1002/bip.360030207.
5
Kinetics of renaturation of DNA.DNA复性动力学
J Mol Biol. 1968 Feb 14;31(3):349-70. doi: 10.1016/0022-2836(68)90414-2.
6
The effects of aqueous neutral-salt solutions on the melting temperatures of deoxyribonucleic acids.中性盐水溶液对脱氧核糖核酸熔点的影响。
Biopolymers. 1971;10(1):47-68. doi: 10.1002/bip.360100106.
7
Kinetics of the helix-coil transition in DNA.DNA中螺旋-线圈转变的动力学
Biopolymers. 1972;11(7):1435-84. doi: 10.1002/bip.1972.360110711.
8
The determination of molecular weight of bacterial genome DNA from renaturation rates.通过复性速率测定细菌基因组DNA的分子量
Eur J Biochem. 1970 Jan;12(1):143-53. doi: 10.1111/j.1432-1033.1970.tb00831.x.
9
Nucleic acid reassociation in formamide.核酸在甲酰胺中的重缔合
Biochemistry. 1969 Aug;8(8):3289-95. doi: 10.1021/bi00836a024.
10
Estimate of the genome size of various microorganisms.各种微生物基因组大小的估计。
J Bacteriol. 1969 Jun;98(3):1400-1. doi: 10.1128/jb.98.3.1400-1401.1969.
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