MLH fusion antibacterial peptide, preparation method and application thereof

A hybrid antimicrobial peptide and antimicrobial peptide technology, applied in the field of genetic engineering, can solve the problems of high cost of chemical synthesis, low antimicrobial peptide, and difficult extraction, etc., and achieve the effects of wide application prospect, improved safety, and simple operation.

Active Publication Date: 2017-10-20
SHAANXI UNIV OF SCI & TECH
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  • Application Information

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Problems solved by technology

Due to the low content of antimicrobial peptides in organisms, direct extraction is difficult and expensive, and it is difficult to achieve mass production; chemical synthesis methods are also rarely used in large-scale production due to high costs

Method used

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  • MLH fusion antibacterial peptide, preparation method and application thereof
  • MLH fusion antibacterial peptide, preparation method and application thereof
  • MLH fusion antibacterial peptide, preparation method and application thereof

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Embodiment Construction

[0032] The present invention provides a method for preparing MLH fusion antimicrobial peptides, using housefly antimicrobial peptides, LL-37 and Helicobacter pylori peptides as parent peptides, using bioinformatics methods to analyze the amino acid sequence of MLH, and according to E. coli codons Preference the gene fragment encoding the hybrid gene MLH, use SOE-PCR technology to synthesize the desired target gene, connect it with the expression vector pET-32a(+), and transform it into the E. coli expression strain to construct the recombinant genetic engineering strain pET- 32a-MLH / BL21(DE3), the protein fusion expression was induced by IPTG, and finally the antibacterial activity of the obtained fusion expressed hybrid peptide was identified. The results showed that the fusion expressed hybrid peptide was effective against Staphylococcus aureus and Bacillus subtilis Both have obvious antibacterial effects.

[0033] See Figure 5 , The present invention discloses a preparation m...

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Abstract

The invention discloses an MLH fusion antibacterial peptide, a preparation method and application thereof. Housefly antibacterial peptide, LL-37 and helicobacter pylori peptide are adopted as the parent peptide, a bioinformatics method is utilized for analysis to obtain an amino acid sequence of MLH, and a gene fragment of the hybrid gene MLH is encoded according to the E.coli codon preference. SOE-PCR technology is employed to synthesize a required target gene, the target gene is connected to the expression vector pET-32a(+), and the obtained recombinant expression vector is transferred into an escherichia coli expression strain to construct the recombinant genetic engineering strain pET-32a-MLH / BL21(DE3), IPTG induced protein fusion expression is carried out, and finally bacteriostatic activity identification is conducted on the obtained fusion expressed hybrid peptide, and the result indicates that the fusion expressed hybrid peptide has significant bacteriostatic effect on staphylococcus aureus and bacillus subtilis.

Description

Technical field [0001] The invention belongs to the technical field of genetic engineering, and specifically relates to an MLH fusion antibacterial peptide and a preparation method and application thereof. Background technique [0002] In recent years, the abuse of antibiotics has become increasingly serious, and more and more bacteria have developed into strains resistant to traditional antibiotics. Antimicrobial peptides have become an ideal substitute for antibiotics due to their stable physical and chemical properties, broad-spectrum antimicrobial properties, and resistance to target strains, and have received extensive attention. Antimicrobial peptides are a kind of small molecular peptides that are widely found in natural organisms, and they are an important part of the body's innate immunity. It has a different target and mechanism of action from antibiotics, has a good antibacterial effect on antibiotic-resistant bacteria, and has broad-spectrum antibacterial properties....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/70A23L33/18A23K20/195A61K38/17A61K38/16A61P31/04
CPCA23K20/195A23L33/18A61K38/00C07K14/195C07K14/43577C07K2319/00C12N15/62C12N15/70C12N2800/22
Inventor 龚国利魏媛
Owner SHAANXI UNIV OF SCI & TECH
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