Figure 5. PCR-based functional assay with degenerate primers (see Fig. 7C for sequences) derived from the proteobacterial tesB genes.

(A) Agarose gel electrophoresis revealed that a proteobacteria-specific tesB gene probe can be used to amplify the corresponding genes of androgen-degrading aerobes such as C. testosteroni. (B) tesB-like PCR products increased only in the testosterone-treated aerobic sewage. The agarose gel images shown in Fig. 5A,B were cropped to show the relevant data only. The full-length gels are present in Fig. S10. (C) The phylogenetic tree of the tesB genes obtained from aerobic sewage incubated with testosterone for 48 hours. Refer to Fig. S6 for the tesB sequences amplified from the aerobic sewage. The sequence of a gene encoding 3,4-DHSA dioxygenase from M. tuberculosis ATCC 25618 served as an outgroup sequence.