微生物および生化学技術ジャーナル

概要

Exploitation of Denaturing Gradient Gel Electrophoresis in Analysis of Microbial Diversity

Shah MP

An application of gradient gel electrophoresis of the PCR-amplified gene fragments were hydrogenase. Comparative analysis of the [NiFe] hydrogenase gene sequences were designed with five different PCR primers. These primers were tested in various combinations on genomic DNA from different hydrogenase-containing and hydrogenase lacking bacteria. For desulfovibrio species appeared to be only one primer pair specific, while others gave positive results with other bacteria. By using this specific pair of primers, we were able to amplify the [NiFe] hydrogenase genes. However, only after could be set DGGE analysis of these PCR products by the number of different species of desulfovibrio in the samples. DGGE analysis of PCR products from different bioreactors demonstrated up to two radicals, wherein at least five distinguishable bands were detected in the microbial mat sample. Because these groups are probably as many desulfovibrio species in these samples, we conclude that the genetic diversity in the species desulfovibrio natural microbial mat is much greater than in experimental bioreactors.