The Establishment And Application Of Biochem-ical Luminescence Detection Method Of Steroid Hormone Detection

Based on the feature that3α-hydroxysteroid dehydrogenase/carbonyl reductase from Comamonas testosteroni is a key enzyme in the degradation of steroid hormones in the environment,the principle that suppressor protein RapA can specific binding with3α-HSD gene promoter and the phenomenon that steroid hormones can make the firefly luciferase luminescence quenching.we use of recombinant technology,P6plasmid as a template to build a control plasmid pUC-4289and put the reporter gene firefly luciferase gene to connected to the3α-HSD gene promoter behind to constructe reporter plasmids pK-3α-LUC. Then transformed into E.coli HB101respectively, we correspondingly construct regulation and reporter cell-free system. Established the detection method of steroid hormones of biochemistry luminescence, luminous intensity and concentrations of steroid hormones were negatively correlated. Optimized its methodology and preliminary applied.Compared to the results of HPLC,the identical results were got with our biosensor method. This method is a sensitive, rapid, easy operation, high throughput, low cost, can detect all known and unknown environment steroid hormones new detection method.In environmental monitoring and food security areas have greater practical value.