重组质粒DNA在热处理过程中的降解/变性规律研究  

The decay/denaturation of plasmid DNA by thermoctreatment

作　　者：李梦南[1] 任大明[2] 王磊[1] 肖伟[2] 郑广宏[1] 

机构地区：[1]同济大学污染控制与资源化研究国家重点实验室,上海200092 [2]复旦大学遗传学研究所遗传工程国家重点实验室,上海200433

出　　处：《环境科学学报》2008年第5期945-950,共6页Acta Scientiae Circumstantiae

基　　金：国家自然科学基金(No.40571145)~~

摘　　要：生物实验室产生的废弃重组基因片段的排放是造成"基因污染"的途径之一.热处理是目前生物实验室处置废弃重组基因片段、核酸等物质的主要手段.以pET-28b质粒为材料,采用定量PCR技术结合电泳和质粒转化等手段分析了重组基因片段在热处理过程中的降解与失活规律,以及不同离子强度对重组质粒热降解的影响.结果显示,100℃热处理过程中质粒pET28b的降解随时间变化较为明显,降解半衰期约为2.7min;处理30min后仍存在具有转化活性的质粒;实验结果表明,100℃热处理后的废弃重组基因片段排入自然生态系统后仍存在基因转移的可能性.Plasmid pET-28b was used as an example to investigate the decay/denaturation of plasmid DNA during thermo-treatment. The DNA integrity was measured using quantitative PCR plus plasmid transformation and electrophoresis technology. The experimental results showed that prolonging the treatment time could increase the decay of plasmid DNA, and the half-life of pET-28b was about 2.7 min during the thermo-treatment process. But after treatment for 30 min, some pET-28b plasmids still had transforming activity. These results indicate that thermo-treatment at 100 ℃ could not decay and inactivate pET-28b completely. When these thermo-treated recombination plasmids are discharged to the eco-system, they may re-nature and transform, thus resulting in gene diffusion.

关 键 词：热处理 质粒DNA 降解/变性 转化 基因扩散 

分 类 号：X592[环境科学与工程—环境工程]