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Biotechnology
Year: 2015  |  Volume: 14  |  Issue: 4  |  Page No.: 194 - 199

Primers and Probe Design for Identification and Analysis of Oil Degradation Bacteria (Bacillus licheniformis) with Quantitative PCR Technology

Hengyu Hu, Dongfeng Zhao and Qiang Zhang    

Abstract: Oil degradation bacteria (Bacillus licheniformis) were isolated and identified by using quantitative PCR-DGGE technology. The Bacillus licheniformis quantity was evaluated by its DNA content and its DNA content was measured by amplification of the DNA target gene. So the specific primers and probe of target gene were designed. Then, the difference was analyzed between using single Bacillus licheniformis bacteria and using Bacillus licheniformis compounded with other bacteria to degrade oil. The results showed that after 60 days cultivation, the oil degradation rate was only 5.6% by using single Bacillus licheniformis bacteria and DNA content of the single bacteria is 1.3×106 copies. When the single bacteria were mixed with other bacteria, the oil degradation rate was 30.7% and DNA content of the Bacillus licheniformis bacteria was 3.3×106. But, when the Bacillus licheniformis bacteria were removed, the oil degradation rate of mix bacteria was 21.3% and the DNA content of Bacillus licheniformis bacteria was 0. It showed the primers and the probe had good specificity. The oil degradation rate and DNA content of single Bacillus licheniformis bacteria were both 0 in the sterilization control group. Compared with traditional technology, the quantitative PCR-DGGE technology could accurately analyze the role of single bacteria in compound bacteria community.

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