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Method for detecting estrogen in environment

An estrogen and environmental detection technology, applied in the field of environmental biology, can solve the problems of cumbersome and difficult environmental estrogen detection, and achieve the effects of good repeatability, simple and convenient operation, and reliable detection results.

Inactive Publication Date: 2011-08-31
SUN YAT SEN UNIV
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  • Abstract
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Problems solved by technology

These methods are cumbersome and difficult to reliably detect environmental estrogens

Method used

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  • Method for detecting estrogen in environment
  • Method for detecting estrogen in environment

Examples

Experimental program
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Embodiment 1

[0022] The promoter sequence of grouper brain aromatase gene, as shown in SEQ ID NO: 1, was cloned by the applicant himself and has been published on the Internet.

[0023] In this example, the pGL3-Basic firefly luciferase reporter gene carrier of Promega Company was used.

[0024] In the present invention, the promoter sequence of the grouper brain aromatase gene is subcloned into the upstream of the firefly luciferase gene of the pGL3-Basic vector, thereby constructing a reporter plasmid named Eccyp19a1b.

[0025] The grouper brain aromatase gene promoter sequence in this example contains an estrogen response element (ERE) and other natural auxiliary elements, so it can specifically respond to estrogen stimulation.

[0026] Transfer U251-MG cells (purchased from China Center for Type Culture Collection) to 24-well plates (10 per well 5 cells), and then use Invitrogen’s Lipofectamine 2000 (1 μL / well) to clone the reporter plasmid (380 ng), goldfish estrogen receptor express...

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PUM

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Abstract

The invention discloses a method for detecting estrogen in the environment, which comprises the following steps of: connecting a luciferase reporter gene vector with a rockfish aromatase gene pituitary specific promoter, constructing a reporter plasmid, co-transfecting a U251-MG cell to the reporter plasmid, a goldfish estrogen acceptor expression plasmid and an actin plasmid, stimulating the transfected U251-MG cell by using a sample to be tested, and detecting estrogen in the sample by detecting the activity of luciferase. The detection method provided by the invention has good repeatability, high sensitivity and reliable detection result, is simple and convenient to operate and can be widely applied to estrogen detection in the environment.

Description

technical field [0001] The invention relates to the field of environmental biotechnology, in particular to a method for detecting estrogen in the environment. Background technique [0002] Environmental hormones are the general term for chemical substances that exist in the environment and can affect human endocrine functions like hormones. Environmental hormones, especially environmental steroid hormones, can be enriched in the natural food chain, difficult to degrade and inactivate, and can seriously damage the normal hormone physiological activities of organisms. The known environmental steroid hormones mainly include: environmental estrogens and environmental androgens. [0003] Environmental steroid hormones are a large class of endocrine disruptors, in which environmental estrogens can bind to estrogen receptors (ER), thereby simulating estrogen activity and exerting estrogen effects. The impact of environmental steroid hormones on human reproductive health has arous...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/66
Inventor 张为民李木张燊姜海燕张利红
Owner SUN YAT SEN UNIV
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