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Ruditapes philippinarum Defensin B gene, its recombinant protein and application

A Philippine clam, gene recombination technology, applied in the fields of application, genetic engineering, plant gene improvement, etc., to achieve a strong inhibitory effect

Inactive Publication Date: 2012-02-29
YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few studies on the antimicrobial peptides and their antibacterial activities of Philippine clams at home and abroad

Method used

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  • Ruditapes philippinarum Defensin B gene, its recombinant protein and application
  • Ruditapes philippinarum Defensin B gene, its recombinant protein and application
  • Ruditapes philippinarum Defensin B gene, its recombinant protein and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The philippine clam defensin B gene has the base sequence shown in SEQ ID No.1.

[0024] The sequence listing SEQ ID No.1 is:

[0025] cataactgac gagtcaacaa gttccaaaaa atgttcgcaa aagcatgctt tgttttgttc

[0026] tttgtgtgta taatggttgg aagtgcttcg gcaaatcctc agaagagatt aacttgtgag

[0027] tttggaggtg tccaggcatg tgccgctcac tgtatcctgt taggttacac tggtggatgg

[0028] tgcgacggtc ataactactg tcattgcaag acgagtggga aaagggaggc ggagacggcc

[0029] tagtgttgaa atatcaagca gtcctgtatc atagaaatct aaaatttatt cagaacaaaa

[0030] agcattagaaaaagaaaaaaaaaaaaaaaaaaa

[0031] (a) Sequential features

[0032] ●Length: 333bp (coding region 31-243bp)

[0033] ●Type: base sequence

[0034] ●Chain type: single chain

[0035] ●Topological structure: linear

[0036] (b) Molecular type: double-stranded DNA

[0037] (c) Assumption: No

[0038] (d) Antisense: no

[0039] (e) Original source: Philippine clam (Venerupis philippinarum)

[0040] (f) Specific name: cDNA

[0041] The cDNA sequence of ...

Embodiment 2

[0043] Preparation of Philippine clam defensin B:

[0044] 1. Extraction of total RNA from clam philippines and purification of mRNA: use Trizol reagent to extract total RNA from the hemolymph of clam philippines infected with Vibrio anguillarum. 50ml, centrifuged at 2000g for 10min, discarded the supernatant, and added 20ml of Disperse the cells with a high-speed disperser, shake vigorously at room temperature for 10s; add 4ml chloroform, shake vigorously for 30s; centrifuge at 10,000g at 4°C for 10min; draw the supernatant into a new centrifuge tube, add an equal volume of pre-cooled isopropanol (about 15ml), placed at -20°C for more than 1 hour; 4°C 10,000g high-speed centrifugation for 10min; carefully discard the supernatant; wash the precipitate with 5ml 70% ethanol; 4°C 10,000g high-speed centrifugation for 10min, carefully discard Supernatant: Vacuum-dry for 5 minutes, dissolve RNA with about 600 μl RNase-free water, and store at -80°C after complete dissolution. Use...

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PUM

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Abstract

The invention relates to antibacterial peptide, more specifically to a Ruditapes philippinarum Defensin B gene, its recombinant protein, a construction method and an application. The Defensin B gene is as shown in a base sequence in a sequence table SEQ ID NO.1. The gene recombinant protein is as shown in an amino acid sequence in a sequence table SEQ ID NO.2. The Defensin B provided by the invention is Ruditapes philippinarum antibacterial peptide, has a strong inhibitory activity of a plurality of marine Gram-positive and negative pathogenic organisms and has application values of being developed into broad-spectrum antimicrobial drugs, genetic breeding, feed additives and the like.

Description

technical field [0001] The invention relates to antibacterial peptides, in particular to a Defensin B gene of Philippine clam and its recombinant protein, as well as its construction method and application. Background technique [0002] Antimicrobial peptide is a kind of small molecular protein with antibacterial activity existing in organisms, the number of amino acids is less than 100, often positively charged, and a class of small peptides with broad-spectrum antibacterial properties. Defensive polypeptide active substance of pathogenicity of exogenous pathogens, has broad-spectrum antibacterial effect, has inhibitory effect on Gram-positive bacteria, Gram-negative bacteria, and fungi, and can also fight against insects and viruses and kill tumors in animals cells without destroying normal cells in the animal body. Antimicrobial peptides generally have no special receptors when they are antibacterial. They directly cause the perforation of the cell membrane through physi...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/435A61K38/17A61P31/04A61P37/04A23K1/16A23L3/3535A23K20/147
Inventor 吴惠丰张林宝赵建民王清
Owner YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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