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A nucleic acid aptamer h10 with targeted inhibition of Vibrio anguillarum and its application

A nucleic acid aptamer, Vibrio eel technology, which can be used in medical preparations containing active ingredients, antibacterial drugs, pharmaceutical formulations, etc., and can solve problems such as hidden dangers, the increase in the number of vibrio infections, and the risk of aquatic product quality and safety. , to achieve the effect of simple chemical synthesis, short screening period and good application prospects

Active Publication Date: 2022-03-22
JIMEI UNIV
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  • Abstract
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AI Technical Summary

Problems solved by technology

At present, with the development of high-density and intensive aquaculture, the number of cultured animals infected with vibriosis is also increasing, which brings huge economic losses to the aquaculture industry.
Due to the serious harm caused by Vibrio eel to aquaculture, a large amount of antibiotics are mainly used to prevent and control Vibrio, which not only leads to the emergence of bacterial resistance, but also brings huge risks and hidden dangers to the quality and safety of aquatic products

Method used

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  • A nucleic acid aptamer h10 with targeted inhibition of Vibrio anguillarum and its application
  • A nucleic acid aptamer h10 with targeted inhibition of Vibrio anguillarum and its application
  • A nucleic acid aptamer h10 with targeted inhibition of Vibrio anguillarum and its application

Examples

Experimental program
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Effect test

Embodiment Construction

[0022] 1 SELEX screening of nucleic acid aptamers

[0023] (1) Bacteria treatment: Take the cultured Vibrio anguillarum in a centrifuge tube, centrifuge at 6000r / min for 5min, discard the supernatant, wash the bacteria 3 times, add sterile culture medium and mix evenly, measure the OD value of the bacteria solution, and then The bacteria content is about 4×10 8 centrifuge at 6000r / min for 5min, discard the supernatant, wash the bacterial pellet three times with 0.9% normal saline, wash the bacterial pellet once with 1×binding buffer, and finally add 100μL of 2×binding buffer hanging.

[0024] (2) Binding: take ssDNA random library, dilute to 3 μmol / L 100 μL with 2× binding buffer, denature in 95°C constant temperature metal bath for 5 minutes, then ice-bath for 10 minutes, then add to the previous bacterial suspension, mix well Combine at 30°C for 2h in a shaker at 100r / min.

[0025] (3) Washing: After the binding is completed, centrifuge at 6000 r / min for 5 min, discard th...

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Abstract

The present invention relates to a nucleic acid aptamer, in particular to a nucleic acid aptamer H10 capable of targeting and inhibiting Vibrio anguillarum, the nucleic acid sequence of which is shown in SEQ.NO.1. And the application of the nucleic acid aptamer H10 in the preparation of Vibrio anguillarum inhibitors. The present invention takes Vibrio anguillaris as the target, and selects the aptamer (H10) screened by Cell-SELEX technology to analyze its antibacterial function. The invention has the functions of short screening period, convenient chemical synthesis, and the ability to replace antibiotics, etc., and has good application prospects. Provide a theoretical basis for further research on inhibitory aptamers.

Description

technical field [0001] The present invention relates to a nucleic acid aptamer, in particular to a nucleic acid aptamer H10 capable of targeting and inhibiting Vibrio anguillarum and its application. Background technique [0002] foreword [0003] Vibrio anguillarum is one of the most common opportunistic pathogens in aquaculture. The aquaculture animal diseases caused by it are prevalent in the world, and can infect more than 50 kinds of sea and freshwater organisms, including rainbow trout, eel, sweetfish, perch, cod, Turbot, flounder, yellow croaker, etc. At present, with the development of high-density and intensive aquaculture, the number of vibriosis in farmed animals is also increasing, which brings huge economic losses to the aquaculture industry. Due to the serious harm caused by Vibrio anguillaris to aquaculture, a large amount of antibiotics are mainly used to prevent and control Vibrio, which not only leads to the emergence of bacterial resistance, but also bri...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/115A61K31/7088A61P31/04
CPCC12N15/115A61P31/04C12N2310/16
Inventor 郑江林筱钧江兴龙鄢庆枇
Owner JIMEI UNIV
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