Departamento de Genética Molecular y Microbiología and Millennium Nucleus on Microbial Ecology and Environmental Microbiology and Biotechnology, Facultad de Ciencias Biológicas, Pontificia Universidad Católica de Chile, Santiago, Chile.
Lett Appl Microbiol. 2010 Jun 1;50(6):552-62. doi: 10.1111/j.1472-765X.2010.02832.x. Epub 2010 Mar 17.
In the last decades, the worldwide increase in copper wastes release by industrial activities like mining has driven environmental metal contents to toxic levels. For this reason, the study of the biological copper-resistance mechanisms in natural environments is important. Therefore, an appropriate molecular tool for the detection and tracking of copper-resistance genes was developed.
In this work, we designed a PCR primer pair to specifically detect copper P-type ATPases gene sequences. These PCR primers were tested in bacterial isolates and metagenomic DNA from intertidal marine environments impacted by copper pollution. As well, T-RFLP fingerprinting of these gene sequences was used to compare the genetic composition of such genes in microbial communities, in normal and copper-polluted coastal environments. New copper P-type ATPases gene sequences were found, and a high degree of change in the genetic composition because of copper exposure was also determined.
This PCR based method is useful to track bacterial copper-resistance gene sequences in the environment.
This study is the first to report the design and use of a PCR primer pair as a molecular marker to track bacterial copper-resistance determinants, providing an excellent tool for long-term analysis of environmental communities exposed to metal pollution.
在过去几十年中，采矿等工业活动导致的铜废物排放量在全球范围内增加，使环境中的金属含量达到了有毒水平。因此，研究自然环境中的生物铜抗性机制非常重要。为此，开发了一种用于检测和跟踪铜抗性基因的适当分子工具。
在这项工作中，我们设计了一对 PCR 引物，用于特异性检测铜 P 型 ATP 酶基因序列。这些 PCR 引物在细菌分离物和受铜污染的潮间带海洋环境的宏基因组 DNA 中进行了测试。此外，还使用 T-RFLP 指纹图谱分析了这些基因序列，以比较正常和铜污染沿海环境中微生物群落中这些基因的遗传组成。发现了新的铜 P 型 ATP 酶基因序列，并确定了由于铜暴露而导致的遗传组成的高度变化。
基于 PCR 的这种方法可用于跟踪环境中的细菌铜抗性基因序列。
这项研究首次报告了设计和使用 PCR 引物对作为分子标记来跟踪细菌铜抗性决定因素，为长期分析暴露于金属污染的环境群落提供了一个极好的工具。
