Pinctada martensii superoxide dismutase (PmECSOD) and application thereof

A technology of pinnacle martensii and superoxide, applied in the field of genetic engineering, can solve problems such as death of pinnacle martensii and economic loss of the pearl industry, and achieve the effect of inhibiting growth

Active Publication Date: 2016-04-13
GUANGDONG OCEAN UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] Pinctadamartensii is one of the main pearl oysters in my country's artificial seawater cultivation. In recent years, due to the deterioration of coastal water qualit

Method used

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  • Pinctada martensii superoxide dismutase (PmECSOD) and application thereof
  • Pinctada martensii superoxide dismutase (PmECSOD) and application thereof
  • Pinctada martensii superoxide dismutase (PmECSOD) and application thereof

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Experimental program
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Effect test

Embodiment 1

[0030] 1. Induction of Pinctada martensii and collection of serum: After the Pinctada martensii was reared for three days, inactivated Escherichia coli and Micrococcus luteus were injected into the adductor muscle of Pinctada martensii for induction, and each injection 10 μL of inactivated Escherichia coli and 20 μL of inactivated Micrococcus luteus were put back into seawater for 12 hours, and then the hemolymph was collected from the adductor muscle of Pinctada martensii with a sterile syringe, centrifuged at 3000rpm for 15min at 4°C, and taken The supernatant was freeze-dried to obtain dry powder of the serum of Pinctada martensii.

[0031] 2. Separation and purification of Pinctada martensii serum: use SunFire TM The semi-preparative C18 column was used to separate and purify the serum of Pinctada martensii, and the results were as follows: figure 1 As shown, four concentration gradients (5% acetonitrile, 18% acetonitrile, 40% acetonitrile and 60% acetonitrile) were elute...

Embodiment 2

[0041] 1. Tissue differential expression analysis of Pinctada martensii PmECSOD: 10 adult Pinctada martensii were used for tissue differential expression analysis. The tissues taken included adductor muscle, mantle, hepatopancreas, gills, gonads and blood cells. Total RNA was extracted and reverse transcribed into cDNA for tissue-specific expression analysis. According to the conserved sequence of Pinctada martensii PmECSOD gene, fluorescent quantitative primers were designed, and the internal reference used was GAPDH. The primers used for fluorescence quantitative analysis are listed in Table 2.

[0042]

[0043] Fluorescence quantitative results such as Figure 11 As shown, the highest expression tissue is gills, which is about 2.9 times that of blood, with a statistical difference (P<0.05); followed by hepatopancreas, which is about 0.29 times that of hemolymph, which also has a significant difference (P<0.05); The mantle and gonad are about 1.15 times and 0.67 times t...

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Abstract

The invention belongs to the technical field of genetic engineering, and particularly discloses pinctada martensii superoxide dismutase (PmECSOD) and an application thereof. The amino acid sequence of the PmECSOD is shown as SEQ ID NO:1; the nucleotide sequence of a PmECSOD gene is shown as SEQ ID NO:2 or SEQ ID NO:3. Compared with traditional SOD, the PmECSOD has many differences and is new SOD through preliminary identification, and the PmECSOD has the effects of inhibiting growth of escherichia coli and growth of micrococcus luteus and has good application prospects.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to superoxide dismutase PmECSOD of Pinctada martensii and its application. Background technique [0002] Pinctadamartensii is one of the main pearl oysters in my country's artificial seawater cultivation. In recent years, due to the deterioration of coastal water quality, various diseases have occurred frequently, resulting in a large number of deaths of Pinctadamartensii due to diseases. The pearl industry has caused huge economic losses. Therefore, understanding the immune mechanism of Pinctada martensii and improving the stress resistance of Pinctada martensii are problems that need to be solved urgently. [0003] Invertebrates do not have the specific immune system of higher animals, so it is very important for the natural immune molecule superoxide dismutase (SOD) to exert immune function in vivo. Superoxide dismutase can scavenge free radicals in the body (O 2- )...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12N15/53C12N15/11C12Q1/68A61P31/04
CPCC12N9/0089C12Q1/6876C12Q2600/158C12Y115/01001Y02A50/30
Inventor 梁海鹰朱家萍雷倩楠吴羽媛王志新
Owner GUANGDONG OCEAN UNIVERSITY
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