Two vectors were used to construct the recombinant gene yeast cell that can be used to bioassay of the pollution of tetracycline antibiotics in the environment. In the expression vector, the GPD (glyceraldehyde-3-phosphate dehydrogenase) promoter was used to drive the gene expression of tetracycline repressor protein (TR) fused with V5 antigen epitope gene, while in the reporter vector, the tetracycline response element (TRE) was used to regulate Lac Z report gene expression. The specificity and the sensitivity of the recombinant gene yeast cell were evaluated respectively by different concentrations of tetracycline antibiotics and non-tetracycline antibiotics. The results showed that there were significant dose effect relationships between the tetracycline antibiotics and the yeast cells, while non-tetracycline antibiotics showed no dose effect relationships with this biosensor. It is illustrated that the recombinant yeast cells can be used to monitor the tetracycline antibiotic pollution on the environment.