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Pinctada martensi lectin PmCLEC-1 and application thereof

A technology of Pinctada martensii and lectin, which is applied in the field of genetic engineering, and can solve problems such as the lack of in-depth reports on the role of lectin

Inactive Publication Date: 2016-05-18
GUANGDONG OCEAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are relatively few studies on the lectin of Pinctada martensii. Hu Yuting cloned two C-type lectin genes, PoLEC-1 and PoLEC-2, from Pinctada martensii, but the lectin gene in Pinctada martensii The role of shellfish in the immune system has not been well reported

Method used

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  • Pinctada martensi lectin PmCLEC-1 and application thereof
  • Pinctada martensi lectin PmCLEC-1 and application thereof
  • Pinctada martensi lectin PmCLEC-1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] 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.

[0032] 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

[0042] 1. Tissue-wide differential expression analysis of Pinctada martensii PmCLEC-1

[0043] In this experiment, GAPDH was used as an internal reference gene, and the results showed that ( Figure 12 ), the expression level in the mantle was the highest, about 128 times that of hemolymph, and the difference was statistically significant (P<0.01), showing a very significant difference; followed by hepatopancreas, about 111 times that of hemolymph; The expression level in the hemolymph is the lowest; the adductor muscle is about 11 times (P<0.05) than the hemolymph; the gills and gonads are 5.9 (P<0.05) times and 10 times (P<0.05) than the hemolymph , fluorescent quantitative primers are shown in Table 2.

[0044]

[0045] 2. Temporal expression of lectin mRNA in hemolymph after bacterial stimulation

[0046] Take 140 healthy Pinctada martensii that have been raised temporarily for 3 days, and divide them into 2 groups: bacterial stimulation group and PBS control group,...

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Abstract

The invention belongs to the technical field of genetic engineering, and particularly discloses pinctada martensi lectin PmCLEC-1 and application thereof. The amino acid sequence of the pinctada martensi lectin PmCLEC-1 is as shown in the SEQ ID NO:1; and the nucleotide sequence of the pinctada martensi lectin PmCLEC-1 is as shown in the SEQ ID NO:2 or 3. Furthermore, the pinctada martensi lectin PmCLEC-1 has an effect of inhibiting growth of colibacillus and micrococcus luteus, and has excellent application prospects.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to the lectin PmCLEC-1 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 lectin (Lectin) to exert immune function in the body. Lectin is ubiquitous in organisms and has a glycosyl binding site. It is a highly sp...

Claims

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Application Information

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IPC IPC(8): C07K14/435C12N15/12C12N15/11C12Q1/68A61P31/04
CPCC07K14/43504C12Q1/6876C12Q2600/158Y02A50/30
Inventor 梁海鹰雷倩楠吴羽媛朱家萍王志新
Owner GUANGDONG OCEAN UNIVERSITY
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