Fluorescent probe for detecting microcystic toxin-LR in water

A microcystin and fluorescent probe technology, applied in the fields of molecular biology and microbiology, can solve the problems of long cycle time, cumbersome preparation of MC-LR monoclonal antibody, and inability to detect on-site

Inactive Publication Date: 2013-05-22
CHANGCHUN UNIV OF SCI & TECH
View PDF7 Cites 11 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The first two methods are either due to high cost, long cycle, and high technical requirements; or due to the complexity of the experimental steps, expensive instruments, time-consuming pretreatment, and inability to detect on-site. Therefore, the main de...

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Fluorescent probe for detecting microcystic toxin-LR in water
  • Fluorescent probe for detecting microcystic toxin-LR in water
  • Fluorescent probe for detecting microcystic toxin-LR in water

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0094] Using MC-LR Standards to Establish Standard Detection Curves for Fluorescent Probes

[0095] 3. Screening of specific affinity ligands (screening includes non-specific elution and specific elution)

[0096] (1) The first round of non-specific elution

[0097] a. Use 0.1mol / L, pH 8.6 NaHCO with the prepared MC-LR 3 The solution was prepared as 100 μg / ml MC-LR solution. Add 1ml of this solution to a sterile polystyrene Petri dish (60×15mm) for coating, and carefully rotate repeatedly until the surface of the Petri dish is completely wet. After completion, put the culture dish into a plastic box covered with wet gauze and incubate at 4°C for more than 12 hours.

[0098] b. After incubation for 12 hours, pour off the coating solution in the petri dish, and vigorously pat on the filter paper that has been irradiated by ultraviolet light to remove the residual coating solution. After removing the residual liquid, add 2ml of blocking solution to the Petri dish, and blo...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a fluorescent probe for detecting microcystic toxin-LR in water, which belongs to the field of molecular biology and microbiology. According to the fluorescent probe, MC-LR is taken as a target, a dodecapeptide library randomly shown on the surface of a bacteriophage is used for selecting a recombination bacteriophage which is combined with the MC-LR in a specific manner, and the single-stranded DNA (Deoxyribonucleic Acid) of the recombination bacteriophage is extracted and recombined for sequencing and sequence alignment, thereby obtaining an MC-LR specific affinity ligand sequence: His-Phe-Phe-Lys-Trp-His-Thr-Arg-Thr-Asn-Asp-Gln, and subsequently, obtaining the specifically combined fluorescent probe FITC-Acp-His-Phe-Phe-Lys-Trp-His-Thr-Arg-Thr-Asn-Asp-Gln through solid-phase polypeptide synthesis and fluorescent marks. The fluorescent probe has the beneficial effects that firstly, a completely new tool is provided for the specific identification and content detection of the MC-LR; and secondly, the fluorescent ligand probe can be used for qualitatively identifying the MC-LR and quantitatively detecting the content of the MC-LR in a sample.

Description

technical field [0001] The invention belongs to the field of molecular biology and microbiology, and relates to screening a ligand sequence with specific affinity for microcystin-LR by using a random display peptide library on the surface of a phage, synthesizing the ligand by artificial solid-phase synthesis, and Fluorescently labeled it to prepare a fluorescent probe for detecting microcystin-LR. Background technique [0002] Microcystins (MCs) are a class of hepatotoxins released by harmful cyanobacteria blooms with strong cancer-promoting effects. Drinking water and aquatic products polluted by MCs pose a huge threat to human health. Among them, Microcystin-LR (Microcystin-LR, MC-LR) is the most toxic of MCs, and it is a biotoxin that poses a potential threat to public drinking water safety. [0003] Studies have shown that MC-LR can cause death in poultry, livestock and wild animals. And low doses of this toxin in drinking water can cause liver damage and promote the ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C09K11/06C07K7/08C07K1/06C07K1/04G01N21/64
Inventor 张淑华于源华廖晓林王晴
Owner CHANGCHUN UNIV OF SCI & TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap