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血清中大肠杆菌染色体DNA和质粒DNA的降解

Degradation of Escherichia coli chromosomal and plasmid DNA in serum.

作者信息

Rozenberg-Arska M, Salters E C, van Strijp J A, Hoekstra W P, Verhoef J

出版信息

J Gen Microbiol. 1984 Jan;130(1):217-22. doi: 10.1099/00221287-130-1-217.

Abstract

Incubation of serum-sensitive [3H]thymidine labelled Escherichia coli PC2166 (RSF1030) and E. coli AM1281 (pBR322) harbouring small plasmids (mol. wt 5.5 X 10(6) and 2.6 X 10(6] in serum resulted in killing of 99.9% of the bacteria within 15 min and in the release of 85% of the radioactivity into the medium after 1 h incubation. The fate of chromosomal and plasmid DNA during incubation of the bacteria in serum was analysed by measurement of the amount of DNA-associated radioactivity, by TCA precipitation, by agarose gel electrophoresis and by the capacity of DNA to transform competent acceptor bacteria. Chromosomal DNA and high molecular weight plasmid DNA were rapidly degraded after 1 h incubation of bacteria in serum. However, low molecular weight plasmid DNA was virtually unaffected and remained physicochemically as well as biologically intact during up to 4 h of incubation of bacteria in serum.

摘要

将血清敏感的、用[3H]胸苷标记的大肠杆菌PC2166(RSF1030)和携带小质粒(分子量分别为5.5×10(6)和2.6×10(6))的大肠杆菌AM1281(pBR322)在血清中培养,15分钟内99.9%的细菌被杀死,培养1小时后85%的放射性物质释放到培养基中。通过测量与DNA相关的放射性物质的量、三氯乙酸沉淀、琼脂糖凝胶电泳以及DNA转化感受态受体细菌的能力,分析了细菌在血清中培养期间染色体DNA和质粒DNA的命运。细菌在血清中培养1小时后,染色体DNA和高分子量质粒DNA迅速降解。然而,低分子量质粒DNA几乎未受影响,在细菌于血清中培养长达4小时的过程中,其物理化学性质和生物学性质均保持完整。

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