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RNA adaptor capable of restraining bacillus typhosus from invading to cells of human body, and preparation method

A typhoid bacillus and human cell technology, applied in the field of microbial infection immunity and inspection, can solve problems such as the lack of perfect typhoid vaccine and high fever

Inactive Publication Date: 2005-01-12
WUHAN UNIV
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AI Technical Summary

Problems solved by technology

Most Salmonella only cause enteritis, but Salmonella typhi can invade human blood vessels and cause high fever and more serious enteric fever and sepsis, which is a difficult problem in clinical treatment
It is a pity that there is no perfect typhoid vaccine in the world so far; and in the past ten years, typhoid bacilli have shown a tendency to be resistant to multiple antibiotics (such as resistance to chloramphenicol, etc.), which has also become a very serious problem

Method used

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Embodiment Construction

[0031] An RNA aptamer capable of inhibiting Salmonella typhi from invading human cells, the RNA sequence of the RNA aptamer is: 5'-GGGAACAGUCCGAGCCUCACUGUUAUCCGAUAGCAGCGCGGGAUGAGGGUCAAUGCGUCAUAGGA-3'.

[0032] A method for preparing an RNA aptamer capable of inhibiting Salmonella typhi from invading human cells is carried out in the following steps:

[0033] 1. Preparation and purification of glutathione sulfur transferase-IVB ciliary structural protein fusion protein-Sepharose 4B complex (GST-PilS-Sepharose 4B). Inoculate the glutathione sulfur transferase-IVB type ciliary structural protein fusion protein recombinant plasmid strain in 2×TY liquid medium with a concentration of 50 μg / ml ampicillin, and culture at 30°C with shaking until the absorbance at 600 nm is 1-2 , add isopropylthioβ-D-galactoside (IPTG) to induce the concentration of 0.1mmol / L, shake culture at 30°C for 4 hours, concentrate the strain, ultrasonically break, add trinitrotoluene-100 (TritonX-100 ) to mak...

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PUM

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Abstract

This invention discloses a RNA adaptor preventing typhoid typhi from intruding into human body cells. This invention discloses a method for preparing the RNA adaptors by preparing and purifying TVB type cilium structure fusion protein-agarose 4B, constructing single strand DNA library randomly, comining double-strand DNA library, amplifying PCR, preparing RNA libraries, screen SELEX, selecting RCR to get RNA adaptors, which can be the agonist of typhi.

Description

Technical field: [0001] The invention belongs to the field of microorganism infection immunity and inspection. It involves the design and preparation of a high-affinity RNA aptamer (Aptamer) that can be used to design and prepare an RNA aptamer (Aptamer) that can bind to the specific pathogenic virulence island type IVB cilium structural protein of Salmonella Typhi that infects humans by using molecular biology technology SELEX technology. Background technique: [0002] Among the nearly 2,300 related Salmonella species, Salmonella typhi is the most important pathogenic bacterium and the only bacterium that is only pathogenic to humans and causes typhoid enteric fever. Salmonella typhi can spread and spread through contaminated food and water. The number of people infected with typhoid bacillus in the world reaches 1.66 billion every year, and typhoid fever is also prevalent in my country for a long time. Recently, my country has designated typhoid bacillus infection to hum...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61P31/04C12N15/10C12N15/115C12N15/62C12P19/34C12Q1/68
Inventor 章晓联潘勤
Owner WUHAN UNIV
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