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通过限制性内切酶切割和相容性测试对SmSu质粒进行表征。

Characterization of SmSu plasmids by restriction endonuclease cleavage and compatibility testing.

作者信息

Grinter N J, Barth P T

出版信息

J Bacteriol. 1976 Oct;128(1):394-400. doi: 10.1128/jb.128.1.394-400.1976.

Abstract

Twelve plasmids carrying genes for streptomycin and sulfonamide resistance were studied for the number and distribution of sites on the plasmid moleucles susceptible to cleavage by the restriction endonuclease EcoRI. Ten of the twelve were found to have a single cut site, one plasmid (R678) had three such sites, and plasmid PB165, which was isolated as three supercoiled deoxyribonucleic acid species with molecular weights 7.4 x 10(6), 14.7 x 10(6), and 21.4 x 10(6) was reduced to a single (linear) species of molecular weight 7.6 x 10(6) after cutting with EcoRI. We conclude that PB165 forms oligomers in Escherichia coli and that the number of copies of these per chromosome is more consistant and that the number of copies of these per chromosome is more consistent with a negative than a positive control mechanism for plasmid replication. Compatibility testing of a positive control mechanism for plasmid replication. Compatibility testing of these plasmids showed they all belong to the same incompatibility group, which we designate IncQ, suggesting that they may have come from a common ancestor.

摘要

对携带链霉素和磺胺抗性基因的12种质粒进行了研究,以确定质粒分子上易被限制性内切酶EcoRI切割的位点数量和分布。发现12种质粒中有10种具有单个切割位点,一种质粒(R678)有三个这样的位点,而质粒PB165,它以分子量分别为7.4×10⁶、14.7×10⁶和21.4×10⁶的三种超螺旋脱氧核糖核酸形式分离得到,在用EcoRI切割后被还原为分子量为7.6×10⁶的单一(线性)形式。我们得出结论,PB165在大肠杆菌中形成寡聚体,并且每条染色体上这些寡聚体的拷贝数更一致,而且每条染色体上这些寡聚体的拷贝数与质粒复制的负调控机制比正调控机制更相符。对这些质粒进行的相容性测试表明它们都属于同一个不相容群,我们将其命名为IncQ,这表明它们可能来自一个共同的祖先。

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