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通过染料浮力密度离心法从细菌裂解物中获得更准确的共价闭合环状质粒与染色体比率的方法。

Method for obtaining more-accurate covalently closed circular plasmid-to-chromosome ratios from bacterial lysates by dye-buoyant density centrifugation.

作者信息

Womble D D, Taylor D P, Rownd R H

出版信息

J Bacteriol. 1977 Apr;130(1):148-53. doi: 10.1128/jb.130.1.148-153.1977.

DOI:10.1128/jb.130.1.148-153.1977
PMID:323223
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC235186/
Abstract

A method that gives high recovery of deoxyribonucleic acid (DNA) from crude bacterial lysates using ethidium bromide-cesium chloride density gradient centrifugation is presented. After Pronase digestion and shearing of the lysate, essentially 100% recovery of chromosomal DNA and a reproducible recovery of covalently closed circular (CCC) plasmid DNA is obtained for a specific plasmid in a given strain. This method should be useful for comparing the CCC plasmid/chromosome ratio of various plasmid-host combinations.

摘要

本文介绍了一种使用溴化乙锭-氯化铯密度梯度离心法从粗制细菌裂解物中高效回收脱氧核糖核酸（DNA）的方法。在蛋白酶K消化和裂解物剪切之后，对于给定菌株中的特定质粒，可实现染色体DNA基本100%的回收率以及共价闭合环状（CCC）质粒DNA可重复的回收率。该方法对于比较各种质粒-宿主组合的CCC质粒/染色体比率应是有用的。

相似文献

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Method for obtaining more-accurate covalently closed circular plasmid-to-chromosome ratios from bacterial lysates by dye-buoyant density centrifugation.通过染料浮力密度离心法从细菌裂解物中获得更准确的共价闭合环状质粒与染色体比率的方法。
J Bacteriol. 1977 Apr;130(1):148-53. doi: 10.1128/jb.130.1.148-153.1977.
2
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引用本文的文献

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本文引用的文献

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Replication of R-factors in Proteus mirabilis: replication under relaxed control.奇异变形杆菌中R因子的复制：松弛控制下的复制
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A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells.一种用于检测和分离闭环双链DNA的染料浮力密度法：HeLa细胞中的闭环DNA
Proc Natl Acad Sci U S A. 1967 May;57(5):1514-21. doi: 10.1073/pnas.57.5.1514.
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Circular DNA.环状DNA
Annu Rev Biochem. 1971;40:899-942. doi: 10.1146/annurev.bi.40.070171.004343.
4
Circular DNA forms of colicinogenic factors E1, E2 and E3 from Escherichia coli.来自大肠杆菌的产大肠杆菌素因子E1、E2和E3的环状DNA形式。
J Mol Biol. 1968 Sep 14;36(2):185-94. doi: 10.1016/0022-2836(68)90374-4.
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Asymmetric distribution of guanine plus thymine between complementary strands of deoxyribonucleic acid of members of the Enterobacteriaceae.肠杆菌科成员脱氧核糖核酸互补链之间鸟嘌呤加胸腺嘧啶的不对称分布。
J Bacteriol. 1971 Apr;106(1):238-42. doi: 10.1128/jb.106.1.238-242.1971.
6
Studies on Escherichia coli sex factors: evidence that covalent circles exist within cells and the general problem of isolation of covalent circles.大肠杆菌性因子的研究：细胞内存在共价环的证据及共价环分离的一般问题。
J Bacteriol. 1971 Mar;105(3):722-7. doi: 10.1128/jb.105.3.722-727.1971.
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Molecular recombination between R-factor deoxyribonucleic acid molecules in Escherichia coli host cells.大肠杆菌宿主细胞中R因子脱氧核糖核酸分子间的分子重组
J Bacteriol. 1970 Jul;103(1):166-77. doi: 10.1128/jb.103.1.166-177.1970.
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Replication of a bacterial plasmid and an episome in Escherichia coli.细菌质粒和附加体在大肠杆菌中的复制。
Biochemistry. 1970 Jan 20;9(2):399-406. doi: 10.1021/bi00804a029.
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Properties of the relaxation complexes of supercoiled deoxyribonucleic acid and protein of the R plasmids R64, R28K, and R6K.R质粒R64、R28K和R6K的超螺旋脱氧核糖核酸与蛋白质的松弛复合物的性质
J Bacteriol. 1974 Oct;120(1):545-8. doi: 10.1128/jb.120.1.545-548.1974.
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Round of replication mutant of a drug resistance factor.耐药因子的一轮复制突变体。
J Bacteriol. 1974 Jun;118(3):855-66. doi: 10.1128/jb.118.3.855-866.1974.