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大肠杆菌K-12和B的中间型修饰突变体对脱氧核糖核酸的修饰

Deoxyribonucleic acid modification by intermediate-type modification mutants of Escherichia coli K-12 and B.

作者信息

Bulkacz J, Roulland-Dussoix D, Boyer H W

出版信息

J Bacteriol. 1975 Dec;124(3):1395-402. doi: 10.1128/jb.124.3.1395-1402.1975.

Abstract

The modification of bacteriophages grown on r-m+/- restriction and modification mutants of Escherichia coli K-12 or B appears to be related to the number of restriction-specific sites in the viral genome. Bacteriophage fd and its mutant U1 fd, which carry two and one B-specific sites, respectively, are not modified in vivo by rB-mB+/- mutant strains. In vitro treatment of fd RF-B+/- deoxyribonucleic acid (DNA) or U1 fd RF-B+/- DNA by endo R-Eco B results in cleavage of the substrate DNA. Lambda bacteriophage, after growth in r-m+/- mutant host strains (lambda-K+/- or lambda-B+/-), is partially protected from in vivo degradation by wild-type homospecific strains. Its efficiency of plating on these strains is approximately 10(-2). However, a hybrid phi80-lambda phage which carries only one K-specific site (sklambda-1) is not modified by rK-mK+/- strains. Labeled DNAs from lambda-B+/- and lambda-K+/- phages were used as substrates for endo R-Eco B and endo R-Eco K nucleases. Zonal centrifugation analysis of the products of the reactions indicate that rK-mK+/- mutants do not protect lambda DNA from in vitro degradation by endo R-Eco K. In contrast, rB-mB+/- mutants appear to partially protect lambda DNA from attack by endo R-Eco B.

摘要

在大肠杆菌K - 12或B的r - m±限制与修饰突变体上生长的噬菌体的修饰似乎与病毒基因组中限制特异性位点的数量有关。噬菌体fd及其突变体U1 fd分别携带两个和一个B特异性位点,它们在体内不会被rB - mB±突变菌株修饰。用内切酶R - Eco B对fd RF - B±脱氧核糖核酸(DNA)或U1 fd RF - B±DNA进行体外处理会导致底物DNA的切割。λ噬菌体在r - m±突变宿主菌株(λ - K±或λ - B±)中生长后,能部分免受野生型同种菌株的体内降解。它在这些菌株上的平板接种效率约为10⁻²。然而,仅携带一个K特异性位点的杂合φ80 - λ噬菌体(skλ - 1)不会被rK - mK±菌株修饰。来自λ - B±和λ - K±噬菌体的标记DNA被用作内切酶R - Eco B和内切酶R - Eco K核酸酶的底物。对反应产物的区带离心分析表明,rK - mK±突变体不能保护λ DNA免受内切酶R - Eco K的体外降解。相反,rB - mB±突变体似乎能部分保护λ DNA免受内切酶R - Eco B的攻击。

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DNA restriction and modification mechanisms in bacteria.细菌中的DNA限制与修饰机制
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The Escherichia coli B restriction endonuclease.大肠杆菌B限制性内切酶
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DNA modification and restriction.DNA修饰与限制
Annu Rev Biochem. 1969;38:467-500. doi: 10.1146/annurev.bi.38.070169.002343.

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