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

A technology of environmental hormones and environmental estrogens, which is applied in the field of detection of environmental hormones in waters based on the vitellogenin gene of the money fish, can solve the problems of no sampling time, inaccurate results, and the inability to accurately estimate the exact range of water pollution concentrations, etc.

Inactive Publication Date: 2017-06-13
SHANGHAI OCEAN UNIV
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  • Abstract
  • Description
  • Claims
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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 fresh...

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

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Embodiment

[0035] In this example, the euryhaline money fish was selected as a living model, and a living environment estrogen detection technology for vitellogenin was established; in particular, it relates to an application in water environment hormone detection based on money fish vitellogenin gene.

[0036] Specific steps are as follows:

[0037] 1) The present invention selects Scatophagus argus, an animal with strong adaptability, as a living model, and uses RACE technology to clone and obtain the full-length cDNA sequence of Scatophagus argus gene; specifically, it is designed by degenerate primer method The primers were used to obtain the partial sequence of the VTGAb of the gold fish vitellogenin, and then the full-length amino acid sequence of the gold fish vitellogenin VTGAb was obtained according to the RACE method. figure 1 It can be seen that the full length of the vitellogenin sequence is about 5000bp, and the designed primers are as follows:

[0038] VTGAb degenerate pri...

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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...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2563/107C12Q2545/114C12Q2521/107
Inventor 张俊彬吴迪吴美琴
Owner SHANGHAI OCEAN UNIV
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