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呼肠孤病毒核糖核酸的纯化与性质

Purification and properties of reovirus ribonucleic acid.

作者信息

Iglewski W J, Franklin R M

出版信息

J Virol. 1967 Apr;1(2):302-7. doi: 10.1128/JVI.1.2.302-307.1967.

Abstract

NaClO(4) was employed in a technique for the rapid extraction of reovirus ribonucleic acid (RNA). The extracted RNA, which was purified in a Cs(2)SO(4) equilibrium density gradient, had a buoyant density of 1.61 g/cm(3) and a sedimentation coefficient of 15S in a 7 to 20% sucrose gradient. It was 90% resistant to ribonuclease in a solution of high ionic strength (0.1 m NaCl). The sensitivity of reovirus RNA to ribonuclease increased with decreasing ionic strength. The thermal denaturation transition of the RNA began at 78 C and was complete at 85 C. The T(m) of the transition was 81 C in 0.01 m tris(hydroxymethyl)aminomethane buffer (pH 7.2) containing 0.001 m ethylenediaminetetraacetate. Thermal denaturation of reovirus RNA resulted in the formation of three ribonuclease-sensitive fractions. Denaturation at 25 C in the presence of dimethyl sulfoxide resulted in the formation of two ribonuclease-sensitive fractions.

摘要

高氯酸钠(NaClO₄)被用于一种快速提取呼肠孤病毒核糖核酸(RNA)的技术中。提取的RNA在硫酸铯(Cs₂SO₄)平衡密度梯度中进行纯化,其浮力密度为1.61 g/cm³,在7%至20%的蔗糖梯度中的沉降系数为15S。在高离子强度(0.1 m NaCl)溶液中,它对核糖核酸酶有90%的抗性。呼肠孤病毒RNA对核糖核酸酶的敏感性随离子强度降低而增加。RNA的热变性转变始于78℃,并在85℃时完成。在含有0.001 m乙二胺四乙酸的0.01 m三(羟甲基)氨基甲烷缓冲液(pH 7.2)中,转变的熔解温度(Tₘ)为81℃。呼肠孤病毒RNA的热变性导致形成三个对核糖核酸酶敏感的组分。在二甲基亚砜存在下于25℃进行变性导致形成两个对核糖核酸酶敏感的组分。

相似文献

本文引用的文献

1
THE SECONDARY STRUCTURE OF REOVIRUS RNA.呼肠孤病毒RNA的二级结构
Proc Natl Acad Sci U S A. 1963 May;49(5):707-14. doi: 10.1073/pnas.49.5.707.
3
ELECTRON MICROSCOPE STUDIES ON REOVIRUS RNA.呼肠孤病毒核糖核酸的电子显微镜研究
Proc Natl Acad Sci U S A. 1964 Dec;52(6):1449-55. doi: 10.1073/pnas.52.6.1449.
5
Effect of NaCIO-4 on bacteriophage: release of DNA and evidence for population heterogeneity.
Virology. 1966 Apr;28(4):742-50. doi: 10.1016/0042-6822(66)90258-3.

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