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Application of silkworm BmcecD1 gene encoding mature peptide in preparation of drugs against colibacillus or pseudomonas aeruginosa

A technology of bmcecd1 and gene encoding is applied in the field of silkworm molecular biology research to achieve commercial value and good antibacterial activity

Inactive Publication Date: 2010-05-12
JINAN UNIVERSITY +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The antibacterial peptide BmcecD1 gene of silkworm Cecropins family has been reported (Yang J, Furukawa S, Sagisaka A, Ishibashi J, Taniai K, Shono T, Yamakawa M. cDNA cloning and gene expression of cecropin D, an antibacterial protein in the silkworm, Bombyx mori , Comp.Biochem.Physiol.B Biochem.Mol.Biol.1999, 122:409-414.) and GenBank accession (NM_001043368), but there is no relevant information about the mature peptide molecule of the gene and the activity research of the mature peptide molecule to report

Method used

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  • Application of silkworm BmcecD1 gene encoding mature peptide in preparation of drugs against colibacillus or pseudomonas aeruginosa
  • Application of silkworm BmcecD1 gene encoding mature peptide in preparation of drugs against colibacillus or pseudomonas aeruginosa

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Example 1 Obtaining the Amino Acid Sequence of the Mature Peptide Encoded by the Bombyx mori BmcecD1 Gene

[0025] The silkworm BmcecD1 gene has been publicly reported, and its nucleotide sequence in GenBank has an accession number of NM_001043368. The inventors annotated and analyzed the gene and obtained 33 amino acid sequences of the gene encoding the mature peptide BmcecD1, such as SEQ ID NO: 1 shown.

Embodiment 2

[0026] Embodiment 2 Construction of recombinant expression vector plasmid, transformant bacterial strain and protein expression

[0027] The silkworm antimicrobial peptide BmcecA2 (mature peptide) molecule, the silkworm antimicrobial peptide BmcecB1 (mature peptide) molecule and the silkworm antimicrobial peptide Cecropin D (mature peptide) molecule, these three have been confirmed to have antibacterial activity, among them, the silkworm antimicrobial peptide BmcecA2 gene Nucleotide sequence, its GenBank accession number is NM_001043997, the nucleotide sequence of the silkworm antimicrobial peptide BmcecB1 gene, its GenBank accession number is 1703262B / AAC60501, the nucleotide sequence of the silkworm antimicrobial peptide Cecropin D gene, its GenBank accession number is BAA31507 .

[0028] The silkworm antimicrobial peptide BmcecA2, the silkworm antimicrobial peptide BmcecB1, the silkworm antimicrobial peptide Cecropin D and the silkworm BmcecD1 gene obtained in Example 1 enc...

Embodiment 3

[0069] Embodiment 3 antibacterial activity

[0070] In this embodiment, the single protein of the silkworm antimicrobial peptide BmcecA2 (bombyx mori antimicrobial peptide BmcecB1, silkworm antimicrobial peptide Cecropin D or silkworm antimicrobial peptide BmcecD1) obtained in Example 2, adopts the conventional agar plate hole diffusion method (plate inhibition zone method) And conventional minimum inhibitory concentration method (MIC), with the following 5 kinds of Gram-negative (G - ), Positive (G + ) bacteria as test bacteria, carry out the test of antibacterial activity, the results are shown in Table 1 and Table 2.

[0071] Bacillus thuringiensis (G + )

[0072] Escherichia coli K 12 D. 31 (Escherichia coli K 12 D. 31 )(G - )

[0073] Pseudomonas aeruginosa (G - )

[0074] Seiiatio maicesceis (G - )

[0075] Ralstonia dolaanacearum (G - )

[0076] Table 1 Diameter of inhibition zone (mm)

[0077] Test bacteria

[0078] In above-mentioned table 1,...

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Abstract

The invention discloses an application of silkworm BmcecD1 gene encoding mature peptide in preparation of drugs against colibacillus or pseudomonas aeruginosa. The amino acid sequence of the silkworm BmcecD1 gene encoding mature peptide is shown in SEQID NO: 1; the minimum inhibition concentration of the silkworm BmcecD1 gene encoding mature peptide to colibacillus is 1.25mu M; and the minimum inhibition concentration of the silkworm BmcecD1 gene encoding mature peptide to pseudomonas aeruginosa is 2.5mu M. The silkworm BmcecD1 gene encoding mature peptide can be used in the drug research and development of human, livestock and poultry, can be used to prevent and cure diseases of human, livestock and poultry caused by the infection of colibacillus or pseudomonas aeruginosa and can also used to prepare the biological product additive and prevent the putrefaction and degeneration of biological products caused by colibacillus or pseudomonas aeruginosa.

Description

technical field [0001] The present invention relates to the research field of silkworm (Bombyx mori) molecular biology, in particular to the research on antimicrobial peptides of the silkworm Cecropins family, in particular to the application of the mature peptide encoded by the antibacterial peptide BmcecD1 gene in the silkworm Cecropins family in the preparation of anti-Escherichia coli or Pseudomonas aeruginosa drugs Background technique [0002] Insects have extremely strong adaptability and defense capabilities, the most important mechanism of which is the non-specific immune system to pathogenic substances and harmful infection factors, insect antibacterial peptides (Antibacterial peptides), also known as insect antimicrobial peptides (Insect Amimicrobial Peptide, AMP), as the main response molecule of the insect humoral non-specific immune response mechanism, has a broad-spectrum and high-efficiency killing effect on bacteria and fungi, and has an inhibitory effect on ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61P31/04
Inventor 黄亚东叶明强邓小娟杨婉莹韩冬丁长才程廷才温硕洋夏庆友曹阳
Owner JINAN UNIVERSITY
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