Figure 3
From: Identification of an operon involved in fluoride resistance in Enterobacter cloacae FRM
Genetic organization and expression of the Is3G fragment. (a) Schematic representation of the Is3G fragment loci. Thick black line, chromosomal DNA; filled arrows, six open reading frames with corresponding gene size; dotted lines, complementary DNA (cDNA) transcribed with gene-specific primers for orf5249 and eno in the 3′ direction. (b) Analyses of reverse transcription (RT)-PCR products (left, cDNA transcribed with gene-specific primers for orf5249; right, cDNA transcribed with gene-specific primers for eno). Lanes: M, 100-base-pair (bp) DNA ladder; 1, PCR positive control using E. cloacae FRM genomic DNA as a template; 2, RT-PCR using E. cloacae FRM cDNA as a template; 3, negative control using E. cloacae FRM RNA as a template; 4, PCR negative control without template. (c) Transcriptional analysis of the genes orf5249, crcB, gpmA, eno, uspA, and ppaC in E. cloacae FRM cultured with 1,000 mg/L fluoride in comparison with culture in the absence of fluoride using quantitative RT-PCR. Transcript levels of the tested genes were normalized to the 16 S rRNA gene. (d) The relative transcription levels of the genes eno, uspA, and ppaC. The 16 S rRNA gene was used as an internal control for normalization.
