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Sinonatrix annularis source antibacterial peptide SAC and application

A technology of red-chained Chinese snake and antibacterial polypeptide is applied to the antibacterial polypeptide SAC of red-chained Chinese snake and its application field, which can solve the problems of human health threat, antibiotic pathogenic microorganism resistance and other problems, and achieves small molecular weight, low hemolytic activity, The effect of simple chemical synthesis methods

Active Publication Date: 2015-09-30
WEIFANG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] In recent years, the large-scale abuse of traditional antibiotics has led to more and more serious drug resistance of pathogenic microorganisms, posing a huge threat to human health.

Method used

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  • Sinonatrix annularis source antibacterial peptide SAC and application
  • Sinonatrix annularis source antibacterial peptide SAC and application
  • Sinonatrix annularis source antibacterial peptide SAC and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Cloning of the coding gene of the antibacterial polypeptide SAC derived from the red chain Chinese snake:

[0017] 1) Extraction of total RNA from the lungs of red-linked Chinese snake snakes:

[0018] ①Take 300 mg of red-linked snake lung tissue, put it into a mortar and add liquid nitrogen to grind it into powder, transfer it to an EP tube, add 1 ml of total RNA extraction reagent (Trizol, product of Lifetech, USA), mix well, and then Centrifuge at 12000 rpm for 10 min at 4°C.

[0019] ② Centrifuge to take the supernatant, add 0.2ml chloroform solution, mix vigorously, let stand at room temperature for 10 minutes, then centrifuge at 4°C, 12000rpm for 10 minutes, and discard the precipitate.

[0020] ③ Add an equal volume of isopropanol to the supernatant, leave it at room temperature for 10 minutes, centrifuge at 4°C and 12,000 rpm for 10 minutes, collect the precipitate, wash it once with 75% (V / V) ethanol, and dry it. The precipitate at the bottom of the tube is th...

Embodiment 2

[0036] Chemical synthesis of antibacterial polypeptide SAC derived from red chain Chinese snake snake:

[0037]Ⅰ. The chemical synthesis method of the antibacterial peptide SAC derived from the red-chain Chinese snake snake: According to the mature peptide amino acid sequence deduced from the gene, its complete sequence was synthesized with an automatic peptide synthesizer (433A, Applied Biosystems), and desalted by HPLC reverse-phase column chromatography.

[0038] Ⅱ. Molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF).

[0039] Ⅲ. The purified antimicrobial polypeptide SAC derived from the red chain Chinese snake snake was identified by high performance liquid chromatography (HPLC), its molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF), and its isoelectric point was determined by isoelectric focusing electrophoresis. The amino acid...

Embodiment 3

[0043] Detection of antibacterial activity of antibacterial polypeptide SAC derived from red chain Chinese snake:

[0044] (1) Pick the test strains stored on the slope and evenly spread them on the MH solid medium (purchased from Qingdao Haibo Biotechnology Co., Ltd.) plate, and place the sterilized 0.5cm diameter filter paper on the medium On the surface, 10 μl of 2 mg / ml antibacterial peptide SAC sample solution dissolved in sterilized deionized water was added dropwise, and incubated upside down at 37°C for 18-20 hours to observe whether the inhibition zone was formed or not. If the sample has antibacterial activity, a clear and transparent bacteriostatic zone will be formed around the filter paper, and the larger the bacteriostatic zone, the stronger the antibacterial activity of the sample.

[0045] (2) Determination of the minimum inhibitory concentration (Minimum Inhibitory Concentration) of antibacterial polypeptide SAC derived from red chain Chinese snake snake (2-fo...

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Abstract

The invention belongs to the technical field of biomedicine, and particularly relates to sinonatrix annularis source antibacterial peptide SAC and application. The sinonatrix annularis source antibacterial peptide SAC is composed of 29 amino acid residues, and is linear chain peptide, the molecular weight is 3569.5 Da, and the isoelectric points are 11.59. The antibacterial peptide SAC has the advantages of being small in molecular weight, simple in chemical synthesis method, efficient in antibacterial action broad spectrum and low in hemolytic performance, and has the wide application prospect. In addition, an encoding gene and application of the SAC are related to.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to an antibacterial polypeptide SAC derived from red chain snake snake and its application. Background technique [0002] In recent years, the large-scale abuse of traditional antibiotics has led to more and more serious problems of drug resistance of pathogenic microorganisms, posing a huge threat to human health. The clinical response to drug-resistant microbial infections is to use new or alternative antibiotics that have not been used against drug-resistant microorganisms, so this requires the continuous development of new antimicrobial drugs. [0003] Antimicrobial peptides are natural small molecular polypeptides encoded by the genes of organisms, and are important molecules in the immune system of organisms. They have direct killing effects on bacteria, fungi, viruses and even protozoa. Antimicrobial peptides have the advantages of small molecular weight, simp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/46C12N15/12A61K38/17A61K8/64A61P31/04A61P31/10A01N47/44A01P1/00A01P3/00
CPCA01N47/44A61K8/64A61K35/583A61K38/00C07K14/46
Inventor 王爱丽
Owner WEIFANG UNIV OF SCI & TECH
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