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通过掺入硫代磷酸酯类似物对体外合成的互补DNA进行核酸酶的选择性保护。

Selective protection of in vitro synthesized cDNA against nucleases by incorporation of phosphorothioate-analogues.

作者信息

Schreiber G, Koch E M, Neubert W J

出版信息

Nucleic Acids Res. 1985 Nov 11;13(21):7663-72. doi: 10.1093/nar/13.21.7663.

DOI:10.1093/nar/13.21.7663
PMID:2415922
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC322078/
Abstract

The conditions for the stepwise synthesis of single- (ss) and double-stranded (ds) cDNA using thio-analogues instead of dNTPs are described in this paper. RNA of paramyxovirus Sendai (strain 6/94) serves as template in these experiments. The increased resistance of this alpha S-modified cDNA against several nucleases, like S1-Nuclease, DNase I, Exonuclease III, snake venom Phosphodiesterase (PDE) and the combination of DNase I and PDE is demonstrated.

摘要

本文描述了使用硫代类似物而非脱氧核糖核苷三磷酸（dNTPs）逐步合成单链（ss）和双链（ds）互补脱氧核糖核酸（cDNA）的条件。在这些实验中，副粘病毒仙台（6/94株）的核糖核酸（RNA）用作模板。结果表明，这种α-S修饰的互补脱氧核糖核酸（cDNA）对几种核酸酶（如S1核酸酶、脱氧核糖核酸酶I、核酸外切酶III、蛇毒磷酸二酯酶（PDE）以及脱氧核糖核酸酶I和PDE的组合）具有更高的抗性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d24/322078/8ee2821f6a4f/nar00315-0133-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d24/322078/8ee2821f6a4f/nar00315-0133-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d24/322078/8ee2821f6a4f/nar00315-0133-a.jpg

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引用本文的文献

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本文引用的文献

1
Studies on polynucleotides. III. Enzymic degradation; substrate specificity and properties of snake venom phosphodiesterase.多核苷酸研究。III. 酶促降解；蛇毒磷酸二酯酶的底物特异性及性质
J Biol Chem. 1959 Aug;234(8):2105-13.
2
Deoxynucleoside [1-thio]triphosphates prevent proofreading during in vitro DNA synthesis.脱氧核苷[1-硫代]三磷酸在体外DNA合成过程中阻止校对。
Proc Natl Acad Sci U S A. 1981 Nov;78(11):6734-8. doi: 10.1073/pnas.78.11.6734.
3
Lifelong persistence of paramyxovirus Sendai-6/94 in C129 mice: detection of a latent viral RNA by hybridization with a cloned genomic cDNA probe.
仙台副粘病毒6/94在C129小鼠中的终身持续存在：通过与克隆的基因组cDNA探针杂交检测潜伏病毒RNA
Virology. 1984 Jul 15;136(1):78-88. doi: 10.1016/0042-6822(84)90249-6.
4
Search for Sendai 6/94 viral RNA in the antigen-free cell line Cl-C-2 isolated from human multiple sclerosis brain tissue.在从人类多发性硬化症脑组织中分离出的无抗原细胞系Cl-C-2中寻找仙台6/94病毒RNA。
Infect Immun. 1983 Aug;41(2):675-82. doi: 10.1128/iai.41.2.675-682.1983.
5
Effect of deoxynucleoside phosphorothioates incorporated in DNA on cleavage by restriction enzymes.掺入DNA中的脱氧核苷硫代磷酸酯对限制酶切割的影响。
J Biol Chem. 1982 Jun 10;257(11):6595-9.
6
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Proc Natl Acad Sci U S A. 1981 Dec;78(12):7350-4. doi: 10.1073/pnas.78.12.7350.
7
Purification and properties of S1 nuclease from Aspergillus.来自曲霉的S1核酸酶的纯化及性质
Methods Enzymol. 1980;65(1):248-55. doi: 10.1016/s0076-6879(80)65034-4.
8
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J Biol Chem. 1982 Aug 10;257(15):8879-84.
9
Purification and further properties of single-strand-specific nuclease from Aspergillus oryzae.米曲霉单链特异性核酸酶的纯化及其他特性
Eur J Biochem. 1973 Feb 15;33(1):192-200. doi: 10.1111/j.1432-1033.1973.tb02669.x.
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