Application of scatophagus argus vitellogenin gene to detection of water area environmental hormones

Abstract

The invention discloses application of a scatophagus argus vitellogenin gene to detection of water area environmental hormones. The application comprises the following steps: obtaining a VTGAb full-length sequence according to a RACE method; sampling and extracting liver RNA (Ribonucleic Acid) after injecting estrogen into scatophagus argus in a gradient manner; detecting expression of VTGAb by adopting an RT real-time quantitative PCR (Polymerase Chain Reaction) method; taking EF1-a as an internal control gene and carrying out absolute quantification on cDNA (complementary Deoxyribonucleic Acid) of liver tissues to establish a detection system of a VTGAb transcriptional level; putting scatophagus argus which is not sexually mature into a water body to be detected and sampling and extracting the liver RNA; detecting expression amount of the VTGAb; and comparing with the detection system, and analyzing the pollution degree of the environmental hormones of a water sample. By adopting the application disclosed by the invention, the pollution condition of the environmental hormones can be measured under the condition that the economic loss is relatively small and the single variable of the scatophagus argus is controlled; and the scatophagus argus vitellogenin gene can be used for the pollution degree of the environmental hormones of water samples of seawater, the mouth of a river and a saline water internal lake.

Description

technical field [0001] The invention belongs to the field of molecular biology and environmental pollution detection, and in particular relates to an application of a money fish vitellogenin gene in water environment hormone detection. Background technique [0002] For the pollution of environmental estrogen, chemical detection methods are mainly used, such as liquid chromatography and mass spectrometry (HPLC-MS). The advantage of these methods is high sensitivity, but the analytical instruments required by these methods are expensive, and the cost is very high, so it is difficult to promote and popularize. In addition, these methods have higher requirements on samples. Environmental samples are different from general samples in that they have the characteristics of a wide variety of compounds, low content, complex sample composition, fluidity and instability. When the concentration of the pollutant to be measured is lower than the detection limit of the existing method or...

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Problems solved by technology

[0006] In addition, the quantitative method used in existing technical detection is relative quantitative, which cannot accurately estimate the exact range of water pollution concentration

At the same time, there are few studies on the simultaneous measurement of seawater and freshwater hormone pollution by live fish in the prior art, and there is no gradient sampling time for estrogen-polluted fish in the prior art to determine the best estrogen detection time point. Quantification after all the genes in the body synthesize protein, the result will be inaccurate

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