Application of mussel GST alpha subtype gene in detecting malachite green-polluting aquatic product

A malachite green and aquatic product technology, applied in the field of detection, can solve the problems of undetectable and missed detection of secondary metabolites, and achieve the effect of improving detection ability and high sensitivity

Active Publication Date: 2019-03-26
INSPECTION & QUARANTINE TECH CENT SHANDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the routine detection of toxic and harmful substances in aquatic products is a variety of chemical methods, but they can only detect some toxic and harmful

Method used

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  • Application of mussel GST alpha subtype gene in detecting malachite green-polluting aquatic product
  • Application of mussel GST alpha subtype gene in detecting malachite green-polluting aquatic product
  • Application of mussel GST alpha subtype gene in detecting malachite green-polluting aquatic product

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0026] Example 1. Primer design and establishment of fluorescence quantitative PCR

[0027] 1. Primer design

[0028] Target gene A: The GST alpha(α) subtype gene sequence of Mytilus galloprovincialis (Mytilus galloprovincialis) was obtained through NCBI (GenBank number: JX485635.1, as shown in SEQ ID No. 9), and the sequence analysis software was used to compare it with the mussel Compare the sequences of other GST genes, design primers for PCR amplification of specific fragments of the target gene in the region with the highest specificity, and use the primer analysis function in Primer Premier 5.0 software to analyze the parameters of the designed primers to determine the best The sequence of the upstream primer RA1 is 5'-ttggccggagtagggttaga-3' (SEQ ID NO. 1), the sequence of the downstream primer RA2 is 5'-ttgaaccatacatgttgtat-3' (SEQ ID NO. 2), and the size of the amplified product is 187 bp.

[0029] Target gene B: The GST mu(μ) subtype gene sequence of Mytilus galloprovincia...

Example Embodiment

[0045] Example 2: Using mussel GST subtype genes to detect mussels polluted by malachite green

[0046] Expose mussels (Mytilus galloprovincialis) to water with different malachite green concentrations of 1, 2.5, 5, 10, 20, 30 and 40 μg / L. After 72 hours, take the mussels that have undergone the above treatments and use malachite green. The enzyme-linked test showed that the mussels treated with the above-mentioned different treatments had contaminated malachite green. Extract the mussel liver tissue RNA from the above-mentioned different treatments and reverse transcription to obtain cDNA. Perform fluorescence quantitative PCR according to the method of step 3 in Example 1 to observe the relative relationship between mussel GST alpha subtype gene, mu subtype gene and sigma3 subtype gene The amount of expression varies with the concentration of malachite green. The results are shown in Table 1.

[0047] Table 1. The relative expression of each subtype gene of mussel GST varies wi...

Example Embodiment

[0050] Example 3 Using mussel GST subtype genes to detect mussels contaminated with deltamethrin

[0051] Expose the mussels (Mytilus galloprovincialis) to the water to be tested with different concentrations of 0.2, 0.5, 1, 2, 4, 6 and 8 mg / L deltamethrin. After 96 hours, take the mussels that have undergone the above treatment, and Chromatographic testing showed that the mussels treated with the above-mentioned different treatments were contaminated with deltamethrin. Extract the mussel liver tissue RNA from the above-mentioned different treatments and reverse transcription to obtain cDNA. Perform fluorescence quantitative PCR according to the method of step 3 in Example 1 to observe the relative relationship between mussel GST alpha subtype gene, mu subtype gene and sigma3 subtype gene The expression level changes with the concentration of deltamethrin. The results are shown in Table 2.

[0052] Table 2. Changes in the relative expression levels of mussel GST subtype genes wit...

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Abstract

The invention discloses application of mussel GST alpha subtype gene in detecting a malachite green-polluting aquatic product, in particular to application of a reagent for detecting a transcriptionallevel of the mussel GST alpha subtype gene in detecting the malachite green-polluting aquatic product. A sequence of the mussel GST alpha subtype gene is as shown by SEQ ID No. 9. The reagent is a specific probe for the mussel GST alpha subtype gene or a specific primer for the mussel GST alpha subtype gene. By adopting the application of the invention, a novel detection method with higher sensitivity is provided for the polluted aquatic product and particularly for the malachite green-polluted mussel, and the food safety detection capacity is effectively improved.

Description

technical field [0001] The invention relates to the field of detection, in particular to the application of mussel GST α subtype gene in detection of aquatic products polluted by malachite green. Background technique [0002] Aquatic products are a general term for animals and plants produced by marine and freshwater fisheries, such as fish, shrimp, crab, etc. my country is not only a big producer of aquatic products, but also a big consumer. The quality and safety of aquatic products is not only related to the life and health of our people, but also affects the development of the aquatic product industry, so it is of great significance to the quality and safety inspection of aquatic products. With the abuse of agricultural and veterinary drugs and the increasing environmental pollution, organisms are continuously exposed to the environment where multiple pollutants cross, resulting in the phenomenon that they suffer from multiple pollutions at the same time. At present, t...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q1/6888C12Q2600/158C12Q2600/166C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 张晓梅张鸿伟崔淑华王培锋梁成珠
Owner INSPECTION & QUARANTINE TECH CENT SHANDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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