Detection Of Polycyclic Aromatic Hydrocarbon (pah) Degradation Genes In Polluted Soil By dna Extraction And Colony Hybridization

摘要

The aim of this study is to detect the DNA sequences for the PAH-dioxygenase gene, the key enzyme of the aerobic catabolic for polycyclic aromatic hydrocarbon degradation, in soil microorganisms by using extraction of total DNA, PCR amplification PAH-dioxygenase sequences, and detection with colony hybridization. To determine whether bacteria containing PAH-dioxygenase gene actually exist in environments contaminated by crude oil or not, the ge-nomic DNA was purified using a genomic DNA purification kit. Purified genomic DNA was then amplified with a thermal cycler, and the PCR products were separeted by agarose gel electrophoresis. Pseudomonas sp. strain ARP 26, which is capable of degrading phenanthrene, was used as target for colony hybridization. It was confirmed that Pseudomonas sp. strain ARP 26 demonstrated DNA sequence homology to the digoxigenin-labeled DNA probes used.