A method for detection of PBDEs in the ocean using the DMRT gene of the four-cornered clam

A four-cornered clam, gene detection technology, applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of small distribution of mussels and oysters, achieve short detection time, reduce pollution, and method sensitive effect

Active Publication Date: 2020-02-04
山东通和海洋科技有限公司
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  • Description
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Problems solved by technology

However, the distribution of mussels and oysters in tidal flat areas lacking attachment bases is very small, and mussels and oysters cannot effectively indicate the pollution of marine sediments

Method used

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  • A method for detection of PBDEs in the ocean using the DMRT gene of the four-cornered clam
  • A method for detection of PBDEs in the ocean using the DMRT gene of the four-cornered clam
  • A method for detection of PBDEs in the ocean using the DMRT gene of the four-cornered clam

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] 1. Domestication of experimental organisms

[0040] Two-year-old male clams were used as experimental organisms, domesticated in laboratory glass tanks for 14 days, the water temperature was 16±1°C, the dissolved oxygen was 7.7-8.3, the pH was 7.9-8.1, and the salinity was 33. Change the water every day, and feed Nitzschia crescentus before changing the water.

[0041] 2. Conduct stress experiment

[0042] Four-cornered clams were transferred to 15 small glass tanks to start the experiment, each small tank was filled with 50L seawater, and 100 four-cornered clams were put in a suitable place to breed.

[0043] A total of 5 experimental groups were set up:

[0044] (1) Non-polluted natural seawater control group;

[0045] (2) dimethyl sulfoxide control group;

[0046] (3) 0.1μg / L tetrabromodiphenyl ether (BDE-47) group;

[0047] (4) 1.0μg / L tetrabromodiphenyl ether (BDE-47) group;

[0048] (5) 10μg / L tetrabromodiphenyl ether (BDE-47) group.

[0049] in:

[0050] ...

Embodiment 2

[0094] 1. Domestication of experimental organisms

[0095] Using two-year-old male clams as experimental organisms, the same method as in Example 1 was used to domesticate two-year-old male clams.

[0096] 2. Preparation of marine sediment

[0097] The clean surface 50mm sediment without PBDEs pollution was collected in the intertidal zone of Yantai, and transferred to the laboratory within 30 minutes after collection. Stir the sediment, pass through a 0.5mm nylon sieve, collect the sediment with a particle size of less than 0.5mm, and then stir evenly to achieve homogenization.

[0098] 3. Sediment spiked with PBDEs

[0099] Prepare BDE-47 mother solution (100mg / 0.1mL) with dimethyl sulfoxide, add standard sediment, and set up 5 experimental groups:

[0100] (1) Blank control (only natural seawater without pollution);

[0101] (2) Solvent control group (DMSO and sediment are mixed according to the volume-to-weight ratio of 1:10000);

[0102] (3) 1mg / Kg dry weight tetrabr...

Embodiment 3

[0151] In order to verify the feasibility and accuracy of the method for detecting the PBDEs in the ocean provided by the present invention, we have collected four-cornered clams, seawater and marine sediment from the tidal flats in the west of Laizhou Bay, and have detected seawater and marine sediment with the method provided by the invention The PBDEs in the samples were compared with the results measured by gas chromatography-mass spectrometry.

[0152] In May 2018, we set up 8 survey stations in the tidal flats on the west coast of Laizhou Bay. The latitude and longitude of these 8 survey stations are shown in Table 1.

[0153] Table 1 Latitude and longitude of survey stations on the west coast of Laizhou Bay

[0154] Station number longitude latitude 1 119°17′6″ 37°40′ 2 119°10′ 37°39′ 3 119°0′ 37°38′ 4 118°57′ 37°32′ 5 118°58′ 37°22′ 6 119°59′ 37°16′ 7 119°4′ 37°15′ 8 119°11′ 37°8′

[0155] We collected ...

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Abstract

The invention discloses a method for detecting PBDEs in ocean by using a mactra veneriformis DMRT gene, which comprises the following steps: 1. taking male mactra veneriformis adults at a sampling point, extracting gonad RNA, and carrying out reverse transcription to synthesize cDNA; 2, amplifying the DMRT gene and a GAPDH gene of any mactra veneriformis sample, and drawing a dissolution curve; 3,detecting the change of a fluorescence signal of SYBRGreen in a PCR reaction process to obtain a Ct value of each PCR reaction, taking the GAPDH gene as an internal reference, a 2<-CT> method is adopted to analyze a result, and calculating the relative expression quantity of the DMRT gene (the relative expression quantity of the DMRT gene is significantly negatively correlated with the PBDEs concentration). The method is lower in detection limit, more sensitive, more environmentally friendly, lower in detection cost, shorter in detection time, good in repeatability and high in reliability, mactra veneriformis is convenient to sample, and early warning of PBDEs pollution is achieved.

Description

technical field [0001] The invention relates to a method for detecting polybrominated diphenyl ethers (PBDEs), in particular to a method for detecting PBDEs in the ocean by using the DMRT gene of the four-cornered clam, and belongs to the technical field of marine PBDEs biological detection. Background technique [0002] Polybrominated diphenyl ethers (PBDEs) are a series of aromatic compounds containing bromine atoms. As the most widely used brominated flame retardants in the world, PBDEs have been widely used in electronic appliances, textiles, rubber, plastics, chemicals and other fields. PBDEs can enter the environment through production, use, and waste of products, accumulate in the ocean through atmospheric deposition, surface runoff, and accumulate in marine sediments and organisms. PBDEs pollution spreads all over the coast of my country. PBDEs pollution has been detected in Bohai Bay, Liaodong Bay, Laizhou Bay, Jiaozhou Bay, Xiamen Sea and Pearl River Estuary. In L...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2561/113C12Q2563/107C12Q2545/114
Inventor 房燕梁欢柳茂生王一奇姚蕾邹强王维忠付建革柯可
Owner 山东通和海洋科技有限公司
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