[Establishment of recombinant Lac Z reporter gene-transformed yeast cells for bioassay of androgen-like compounds in environment].

OBJECTIVE

To establish the yeast-based bioassay system for the androgenic endocrine disruptors in the environment.

METHODS

The recombinant gene-transformed yeast cells were constructed based on two episomal vectors. In the expression vector, the expression of androgen receptor (AR) gene was driven by 3-glyceraldehydephosphate dehydrogenase (GPD) promoter and the gene was fused to V5 epitope; in the reporter vector, the expression of the Lac Z gene as a report gene was under the control of the androgen response element (ARE). The vectors were transformed into yeast cells (W303-1A) to construct the yeast-based bioassay system. The sensitivity of this assay system was evaluated by the androgenic compounds (dihydrotestosterone, DHT and testosterone propionate, TP)and the specificity of this system by the estrogenic compounds (17β-estradiol, estriol, estrone, diethylstilbestrol and ethinyl estradiol).

RESULTS

Both DHT and TP yielded a significant dose-effect model with the recombinant gene-transformed yeast cells, indicating that this yeast bioassay system has a high sensitivity. Moreover, the assay also exhibited a high specificity in that the yeast cells had no dose-effect relationship with the different concentrations of five estrogens (17β-estradiol, estriol, estrone, diethylstilbestrol and ethinyl estradiol).

CONCLUSION

The recombinant yeast cells could be applied to screen for the androgen-like compounds in the environment.