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热损伤DNA的存活及生物活性

Survival and biological activity of heat damaged DNA.

作者信息

Masters C I, Miles C A, Mackey B M

机构信息

Institute of Food Research, Reading, Earley Gate, UK.

出版信息

Lett Appl Microbiol. 1998 Nov;27(5):279-82.

PMID:9830145
Abstract

The thermal degradation of plasmid pUC18 held at temperatures between 100 and 135 degrees C was examined by measuring the ability of heat-treated plasmid preparations to transform Escherichia coli to ampicillin resistance using electroporation. Substantial protection against loss of transforming ability during heating was provided by concentrations of NaCl between 0.25 and 2.0 mol l-1. For example, the addition of 1.0 mol l-1 NaCl to samples heated at 100 degrees C for 15 min increased transformation frequency about 200-fold compared with samples heated without NaCl. In the presence of 0.5-2.0 mol l-1 NaCl, transforming capacity was not destroyed even by heating at 121 degrees C for 15 min, i.e. after a typical sterilization treatment. These findings may have implications for the safe disposal of genetically modified micro-organisms and recombinant DNA preparations.

摘要

通过测量经热处理的质粒制剂利用电穿孔法将大肠杆菌转化为氨苄青霉素抗性的能力,研究了在100至135摄氏度之间保存的质粒pUC18的热降解情况。0.25至2.0 mol l-1的NaCl浓度对加热过程中转化能力的丧失提供了显著保护。例如,向在100摄氏度加热15分钟的样品中添加1.0 mol l-1 NaCl,与未添加NaCl加热的样品相比,转化频率提高了约200倍。在存在0.5至2.0 mol l-1 NaCl的情况下,即使在121摄氏度加热15分钟(即经过典型的灭菌处理),转化能力也不会被破坏。这些发现可能对转基因微生物和重组DNA制剂的安全处置具有重要意义。

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