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One group of aptamers for specifically recognizing staphylococcus aureus enterotoxin C1

A technology of staphylococcus gut, nucleic acid aptamer, applied in recombinant DNA technology, microbial determination/inspection, biochemical equipment and methods, etc., can solve the problems of unutilized, expensive, difficult to obtain preparation cost of SEC1 antibody, etc., To achieve the effects of convenient operation, low cost and simple preparation method

Active Publication Date: 2013-08-14
JIANGNAN UNIV
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AI Technical Summary

Problems solved by technology

[0004] At present, the existing domestic and foreign reports on nucleic acid aptamers of protein targets screened by SELEX technology mainly include thrombin, IgE, HIV-1 reverse transcriptase, neuropeptide Y, ricin, botulinum toxin, prostate Specific antigens, DNA-dealkylation enzymes, Staphylococcus aureus enterotoxin B, etc., there is no report on the nucleic acid aptamer sequence of Staphylococcus aureus enterotoxin C1
However, SEC1 antibodies are difficult to obtain and expensive to prepare, and are subject to many conditions when used. Aptamers have more significant advantages over antibodies, but their advantages have not been utilized in research in this area, so related research work needs to be strengthened urgently

Method used

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  • One group of aptamers for specifically recognizing staphylococcus aureus enterotoxin C1
  • One group of aptamers for specifically recognizing staphylococcus aureus enterotoxin C1
  • One group of aptamers for specifically recognizing staphylococcus aureus enterotoxin C1

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Embodiment Construction

[0013] The inventors of this case have found through long-term and practice that the interaction between single-stranded nucleic acid bases can make it form many possible spatial structures, including hairpins, pseudoknots, convex loops, and G-quadruplexes. Various target molecules undergo conformational binding. Based on this conformational recognition, a sufficient capacity (10 14 -10 15 ) nucleic acid library can contain as much three-dimensional structure as possible, so it is possible to screen high-affinity aptamers of any kind of target substance therefrom. Using SELEX technology, the target toxin SEC1 has an exponential evolutionary enrichment effect on each round of screening nucleic acid library. After dozens of rounds of repeated screening, reverse screening and negative screening are combined, and finally a nucleic acid aptamer sequence with high affinity and good specificity is obtained. .

[0014] Based on the above findings, the inventors of the present case ...

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Abstract

The invention discloses one group of aptamers for specifically recognizing staphylococcus aureus enterotoxin C1, and belongs to the biological technical field of food safety. In order to overcome the defect of lacking aptamer of staphylococcus aureus enterotoxin C1 in the prior art, one group of aptamers having good affinity and specificity on the staphylococcus aureus enterotoxin C1 is obtained by 12 turns of repeated screening through SELEX (systematic evolution of ligands by exponential enrichment) technology and analysis and verification of affinity and specificity tests. The group of aptamers provides a specific and efficient recognition ligand for analyzing and detecting the staphylococcus aureus enterotoxin C1 in food or environment and provides a new selection for the existing method of detecting the staphylococcus aureus enterotoxin C1 through antibodies.

Description

technical field [0001] The invention relates to the field of food safety biotechnology, specifically refers to the preparation of a group of high specificity and high specificity of Staphylococcus aureus enterotoxin C1 by using SELEX technology (exponential enrichment ligand system evolution technology) combined with molecular biology and in vitro chemical synthesis Affinity nucleic acid aptamers provide scientific basis and theoretical basis for the development of new detection technology based on nucleic acid aptamer recognition-Staphylococcus aureus enterotoxin C1. Background technique [0002] Staphylococcus aureus Enterotoxins (SEs for short) are a group of small molecular weight (26-30kDa) heat-resistant protein toxins secreted by Staphylococcus aureus outside the cell. Food poisoning caused by this enterotoxin accounts for a very high proportion of bacterial food poisoning and is a worldwide public health problem, of which more than 75% are caused by Staphylococcus au...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/115C12Q1/68C07K14/31
Inventor 王周平黄玉坤夏雨陈秀娟
Owner JIANGNAN UNIV
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