| Estuaries are vital to the ecosystem as they house species of plants, animals, fungi, and bacteria. However, heavy metals are introduced to estuaries anthropogenically, from automobile exhausts, factory emissions, road work material, and natural decaying rocks. In turn, microbes have developed genes that allow them to be resistant to heavy metals. This study was done to detect genes that confer resistance to cadmium, zinc, and lead in Chloroflexi -like microbes. Primers were designed from phylogenetic trees constructed of microbial genomes known to contain these resistant genes. These primers were utilized for amplifying on genomic DNA extracted from environmental samples using standard and nested PCR. The original primers designed did not amplify genes from DNA extracted from estuary samples. However, primers that were designed exactly to Chloroflexus aurantiacus J10-fl worked. While the zntA/cadA/pbrA gene was not identified within the estuary, techniques were developed to optimize the PCR results. For future research, it is highly recommended that fresh soil sediment be obtained, primers be redesigned, and PCR machine parameters be revisited for successful PCR amplification. |
