Transcriptional Analysis of OPH-like Genes

To identify the potential function of the OPH-like genes, total RNA of CF-BD cultivated for 20 h in the BHI medium containing 0, 12.5, 25, and 50 mg trichlorphon were extracted to analyze the expression of the OPH-like genes. Complementary DNA (cDNA) was reverse-transcribed from 2 μg total RNA using MMLV reverse transcriptase (Promega). The recA gene was used as the reference gene [66 (link)]. Real-time quantitative PCR amplification was performed using Mx3000P spectrofluorometric thermal cycler (Agilent Technologies, Santa Clara, CA, USA) and Real Master Mix (SYBR Green) kit (Tiangen), starting with a 2 min incubation at 95 °C, followed by 40 cycles of 95 °C, 20 s; 55 °C, 1 min; and 72 °C, 30 s. Primer information for the genes was described in Additional file 3: Table S3.