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Application of gyra gene pna combined with penetrating peptide combined with antibiotics in the preparation of drugs for inhibiting Riemerella anatipestifer

A technology of Riemeria anatipestifer and membrane-penetrating peptides, which can be applied to antibacterial drugs, medical preparations containing active ingredients, and pharmaceutical formulas, and can solve the problems of unreported antibacterial effects

Active Publication Date: 2020-09-15
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it has not been reported that gyrA gene PNA combined with penetrating peptide combined with antibiotics can improve the antibacterial effect of RA

Method used

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  • Application of gyra gene pna combined with penetrating peptide combined with antibiotics in the preparation of drugs for inhibiting Riemerella anatipestifer
  • Application of gyra gene pna combined with penetrating peptide combined with antibiotics in the preparation of drugs for inhibiting Riemerella anatipestifer
  • Application of gyra gene pna combined with penetrating peptide combined with antibiotics in the preparation of drugs for inhibiting Riemerella anatipestifer

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Example 1, PNA design with RA gyrA as target gene

[0024] The nucleotide sequence of the gyrA gene of RA was retrieved in GenBank (Genbank: CP003787.1), and the similarity analysis of the retrieved gyrA gene among RA strains was performed using BLAST software. A PNA sequence with better binding stability was designed, named PNA-gyrA, and its nucleic acid sequence was 5'-cggttgcccactcc-3' (SEQ ID NO.1). And modify the PNA-gyrA sequence, in order to increase its cell permeability and solubility, add penetrating peptide (CPP)【(KFF) 3 K] and the solubility promoting factor O-Linker, the amino acid sequence of the penetrating peptide is as follows: KFFKFFKFFK (SEQ ID NO.2), synthesized by Panagene, and the synthetic sequence of PNA (CPP-PNA-gyr) combined with the penetrating peptide is as follows: 5 '-FITC-OO-KFFKFFKFFK-OO-CGGTTGCCCACTCC-3', FITC is a fluorescent label for positioning after PNA action, K is lysine, F is phenylalanine, O is glycine, OO is used for connectio...

Embodiment 2

[0025] Embodiment 2, CPP-PNA-gyrA research on the antibacterial effect of Riemerella anatipestifer

[0026] Referring to the operation standard of antimicrobial susceptibility test, the MIC of CPP-PNA-gyrA to the tested strains RACH-1 strain, RA CH-2 strain, and ATCC 11845 strain was detected by microdilution method. The specific steps are as follows:

[0027] (1) Bacterial suspension preparation

[0028] Strain RACH-1, RA CH-2 and ATCC 11845 were inoculated on the TSA agar plate by streaking, and incubated overnight in a 37°C incubator. On the next day, a single colony was picked with an inoculation loop and inoculated in 3 mL of nutrient broth. 37°C, 220r / min constant temperature shaker culture, until the logarithmic growth phase (OD 600 =0.8), the bacterial solution was diluted with TSB to a bacterial content of 2×10 5 CFU / mL.

[0029] (2) Dilution of CPP-PNA-gyrA and antibacterial drugs

[0030] Dilute CPP-PNA-gyrA to 50 μM with sterile water and divide it for use; ofl...

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Abstract

The invention relates to application of cell-penetrating peptide bound gyrA gene PNA (CPP-PNA-gyrA) combined antibiotics in preparation of a medicine for inhibiting riemerella anatipestifer. After theCPP-PNA-gyrA and antibiotics co-act on the riemerella anatipestifer, the MIC (Minimal Inhibitory Concentration) on the riemerella anatipestifer is halved, so that that the CPP-PNA-gyrA is capable ofenhancing the antibacterial effects of the antibiotics. Therefore, the CPP-PNA-gyrA can be compounded with the antibiotics to prepare the medicine for resisting the riemerella anatipestifer.

Description

technical field [0001] The invention belongs to the field of poultry prevention and control, and relates to the application of gyrA gene PNA combined with penetrating peptide combined with antibiotics in the preparation of drugs for inhibiting Riemerella anatipestifer. Background technique [0002] Riemerella anatipestifer (RA) mainly causes contact infectious diseases of ducks, geese, turkeys and other poultry. It is a Gram-negative rod-shaped bacterium with no flagella and no spores. Ducks are the most susceptible and are distributed worldwide, with high morbidity and mortality, which seriously endangers the development of duck industry. At present, the prevention and control of Riemerella anatipestifer is on the one hand through strengthening the feeding management and establishing a perfect disinfection system; on the other hand, antibiotics are used to prevent and control the disease. medicine problem. Therefore, it is necessary to provide a control method for solving...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/711A61K31/4704A61K31/496A61P31/04
CPCA61K31/4704A61K31/496A61K31/711A61P31/04A61K2300/00Y02A50/30
Inventor 程安春吴莉萍邱浩汪铭书
Owner SICHUAN AGRI UNIV
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