Antibacterial peptide His6K and application thereof

An anti-microbial and fusion peptide technology, applied to the antibacterial peptide His6K and its application fields, can solve the problems of low content of natural antibacterial peptides, difficulty in extraction, and high cost of chemically synthesized antibacterial peptides, and achieve the effect of good antibacterial activity and broad application prospects.

Active Publication Date: 2015-11-04
GUANGDONG HINAPHARM PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Antimicrobial peptides have extremely broad application prospects, but due to the low content of natural antimicrobial peptides in organisms, the extraction is difficult, and the cost of chemically synthesizing antimicrobial peptides is too high, it is difficult to use them in large-scale industries such as food and feed

Method used

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  • Antibacterial peptide His6K and application thereof
  • Antibacterial peptide His6K and application thereof
  • Antibacterial peptide His6K and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1, artificial synthesis of antibacterial peptide and antibacterial activity assay

[0050] 1. Synthesis of antimicrobial peptides

[0051] The amino acid sequence of the antimicrobial peptide in the prior art is shown in sequence 1 of the sequence listing. The antimicrobial peptide shown in Sequence 1 of the Sequence Listing was artificially synthesized and named as the antimicrobial peptide Histonin.

[0052] The amino acid sequence of the antimicrobial peptide provided by the present invention is shown in sequence 2 of the sequence listing, and its coding gene is shown in sequence 3 of the sequence listing. The antimicrobial peptide shown in Sequence 2 of the Sequence Listing was artificially synthesized and named as the antimicrobial peptide His6K.

[0053] Second, the determination of the minimum inhibitory concentration MIC of antimicrobial peptides

[0054] Detect the minimum inhibitory concentration MIC of antimicrobial peptides (antimicrobial peptide...

Embodiment 2

[0061] Embodiment 2, the preparation of antimicrobial peptide His6K and antibacterial activity assay

[0062] 1. Construction of recombinant plasmids

[0063] 1. Synthesize the double-stranded DNA molecule shown in the sequence 4 of the sequence listing from the 244-746th nucleotide at the 5' end.

[0064] 2. Double-digest the double-stranded DNA molecule obtained in step 1 with restriction endonucleases XhoI and NotI, and recover the digested product.

[0065] 3. Digest the yeast expression vector pPic9K with restriction endonucleases XhoI and NotI, and recover the vector skeleton of about 9.3 kb.

[0066] 4. Ligate the digested product obtained in step 2 with the vector backbone obtained in step 3 to obtain the recombinant plasmid pPic9K-His6K ( figure 1 ). For the results of enzyme digestion and identification of the recombinant plasmid pPic9K-His6K, see figure 2 ( figure 2 Among them, M is DNA molecular weight marker; Lane 1 is the result of restriction endonuclease...

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Abstract

The invention discloses an antibacterial peptide His6K and application thereof. The antibacterial peptide His6K provided by the invention is the polypeptide shown as the following a) or b) or c): a) a polypeptide with an amino acid sequence as shown in sequence 2 in the sequence table; b) a polypeptide containing more than two single-copies, and the single copy is as shown in the sequence 2 in the sequence table; and c) a fusion polypeptide containing the a) or b). Experiments prove that the antibacterial peptide His6K provided by the invention has good antibacterial activity, which is 1.5 times that of the antibacterial peptide Histonin.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to antibacterial peptide His6K and its application. Background technique [0002] Antibacterial peptides (Antibacterial peptides) are a class of polypeptides that are encoded by specific genes of various biological cells and induced by external conditions, and have broad-spectrum anti-bacteria, fungi, viruses, protozoa, and anti-tumor cells. In 1974, Swedish scientist G.Boman and others injected Escherichia coli into the pupa of Samia Cynthia, and found a basic polypeptide substance with antibacterial activity in blood lymphocytes. After years of research, more than 2,500 antimicrobial peptides have been discovered from animals, plants, bacteria and viruses, and thousands of analog peptides have been artificially synthesized using natural antimicrobial peptides as templates. Antimicrobial peptides have antibacterial, fungal, tumor and wound healing effects, and are an impor...

Claims

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Application Information

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IPC IPC(8): C07K14/00C12N15/11C12N15/81C12N1/19A61K38/16A61P31/04
CPCA61K38/16C07K14/00
Inventor 周玉岩罗科张珂卿逯佩凤
Owner GUANGDONG HINAPHARM PHARMA CO LTD
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