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Metagenomic DNA (metagenomic + dna)
Selected AbstractsIdentifying Natural Product Biosynthetic Genes from a Soil Metagenome by Using T7 Phage SelectionCHEMBIOCHEM, Issue 16 2009Keya Zhang Doing the dirty work: Metagenomic DNA was directly isolated from soil bacteria to construct a T7 phage library. Iterative phage selection by protein modification catalyzed by phosphopantetheinyl transferase enriched putative acyl carrier protein and peptidyl carrier protein genes that are associated with natural product biosynthetic enzymes. [source] Novel polymerase chain reaction primers for the specific detection of bacterial copper P-type ATPases gene sequences in environmental isolates and metagenomic DNALETTERS IN APPLIED MICROBIOLOGY, Issue 6 2010R. De la Iglesia Abstract Aims:, 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. Methods and Results:, 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. Conclusions:, This PCR based method is useful to track bacterial copper-resistance gene sequences in the environment. Significance and Impact of the Study:, 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. [source] Optimization of metagenomic DNA extraction from activated sludge samplesASIA-PACIFIC JOURNAL OF CHEMICAL ENGINEERING, Issue 5 2009Yuan-Yuan Qu Abstract Metagenomic DNA extraction is essential for metagenomic technology. Therefore, optimization of a conventional total DNA extraction from activated sludge was investigated in detail in this study. Throughout two distinct orthogonal experiments, it was shown that the highest yield for metagenomic DNA could be obtained using TENP buffer, lysozyme of 1 mg ml,1 (1 h), protease K (200 µg ml,1), SDS (1%, 1 h). Furthermore, the quality of the differentially extracted DNA was subsequently assessed by the molecular fingerprint technology, such as denaturing gradient gel electrophoresis (DGGE) and ribosomal intergenic spacer analysis (RISA). The results indicated that the microbial diversity was dramatically different by different combined methods, and the DNA template quality for RISA was much better than that for polymerase chain reaction (PCR)-DGGE. This study provides detail process for metagenomic DNA extraction of activated sludge, and it would be useful for metagenomic DNA extraction of other environment samples. Copyright © 2009 Curtin University of Technology and John Wiley & Sons, Ltd. [source] | ||||||||||