快速检测四环素类抗生素的重组Lac Z基因酵母细胞的建立

Construction of a Recombinant Lac Z Gene in Yeast Cell for Rapid Detection of Tetracycline Antibiotics

以载体双表达的方式构建监测环境中四环素类抗生素污染的重组酵母细胞。在表达载体中,用3-磷酸甘油醛脱氢酶(GPD)启动子驱动四环素阻遏蛋白基因(TR)的表达,并使其与V5抗原表位编码基因相融合。在报告载体中,用四环素效应元件(TRE)调控的Lac Z作为报告基因。将两者转化于酵母细胞中,构建成四环素类抗生素调控的重组Lac Z基因酵母细胞。用不同浓度的四环素类抗生素和非四环素类抗生素对重组酵母细胞分别进行敏感度和特异度研究。结果表明四环素类抗生素与该重组酵母细胞报告基因表达水平有明显的剂量效应关系,说明对检测四环素类抗生素有较好的敏感性,而非四环素类药物与重组酵母细胞报告基因表达水平之间无明显的剂量效应关系,说明其检测具有良好的特异性。该重组基因酵母细胞可用于对环境四环素类抗生素污染程度的监测。

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 （glyeeraldehyde-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.