Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Expression protein for detecting bovine viral diarrhea virus antibody, ELISA kit and preparation method and application thereof

A technology of bovine viral diarrhea and kits, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of being unsuitable for use in pastoral areas, time-consuming and labor-intensive, etc., to avoid refolding difficulties, save labor and time, and have wide application prospects Effect

Active Publication Date: 2019-12-10
内蒙古元山生物科技有限公司
View PDF6 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Virus isolation, serum neutralization experiments, and other methods are time-consuming and laborious. RT-PCR, electron microscope observation, etc. require professionals, and are not suitable for use in pastoral areas

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
  • Expression protein for detecting bovine viral diarrhea virus antibody, ELISA kit and preparation method and application thereof
  • Expression protein for detecting bovine viral diarrhea virus antibody, ELISA kit and preparation method and application thereof
  • Expression protein for detecting bovine viral diarrhea virus antibody, ELISA kit and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0059] Optionally, the method for preparing the expressed protein at least includes the following steps:

[0060] (1) Query the E2 gene sequence of bovine viral diarrhea virus, optimize the E2 gene sequence according to the preferred codon table of Escherichia coli, increase the signal peptide sequence and synthesize it, and obtain the optimized SP-E2 gene sequence, SP-E2 gene sequence The nucleotide sequence is shown in SEQ ID NO: 1;

[0061] (2) Construction of the E2 gene expression strain: the nucleotide sequence of the optimized SP-E2 gene was connected to the prokaryotic expression vector pET-32a(+) to obtain the recombinant plasmid pET-32a-SP-E2, and the constructed recombinant plasmid pET-32a-SP-E2 was transformed into Escherichia coli BL21 cells, and the recombinant expression strain of SP-E2 gene was constructed;

[0062] (3) Induced expression and purification of the expressed protein: IPTG was used to induce the expression of the target protein, and the target pro...

Embodiment 1

[0083] An ELISA kit for detecting bovine viral diarrhea virus, its composition is as shown in table 1:

[0084] Table 1

[0085] name composition Quantity (marked amount) sample diluent 0.5 wt% PEG 6000 1 bottle (250ml) 10× Concentrated Washing Solution PBST containing 0.5% (v / v) Tween 20 1 bottle (250ml) Enzyme-labeled antibody HRP-labeled rabbit anti-bovine IgG 1 bottle (30ml) Substrate chromogenic solution TMB 1 bottle (30ml) stop solution 2mol / L dilute sulfuric acid 1 bottle (30ml) 96-well plate coated with BVDV antigen — 5 pieces positive control standard positive serum 1 bottle (2ml) negative control standard negative serum 1 bottle (2ml) Sealing film — 10 sheets manual — 1 copy

Embodiment 2

[0086] Embodiment 2: BVDV-E2 gene bioinformatics analysis and expression

[0087] 1 Materials and methods

[0088] 1.1 Test material

[0089] Escherichia coli BL21 was purchased from Quanshijin Company, DNA restriction endonuclease was purchased from NEB Company, DNA gel recovery and purification kit was purchased from AXYGENG Company, plasmid mini-extraction kit was purchased from QIAGEN Company; yeast powder, tryptone, agar The powder was the product of Oxoid Company, the rabbit anti-bovine IgG labeled with horseradish peroxidase was purchased from Sigma Co., Ltd., and the BCA protein quantitative detection kit was purchased from TAKARA Company.

[0090] 1.2 Test method

[0091] 1.2.1 Bioinformatics analysis of E2 gene

[0092] Using DNAstar, Swiss and other software and online servers to analyze the BVDV (DQ088995.2) E2 gene, and predict the size, signal peptide, secondary structure, tertiary structure, antigenicity, and hydrophobicity of the protein encoded by the gene. ...

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

PropertyMeasurementUnit
molecular weightaaaaaaaaaa
Login to View More

Abstract

The invention relates to the field of biological products, in particular to an expression protein for detecting a bovine viral diarrhea virus antibody, an ELISA kit and a preparation method and application thereof. The expression protein for detecting the bovine viral diarrhea virus antibody is characterized in that a nucleotide sequence for encoding the expression protein is shown as SEQ ID NO: 1, and the expression protein is a supernatant protein. The invention also relates to the ELISA kit for detecting the bovine viral diarrhea virus antibody. The ELISA kit comprises an ELISA plate coatedwith the expression protein. The kit disclosed by the invention has the technical advantages of high biological safety, low cost, high coincidence rate, good repeatability and high specificity and sensitivity.

Description

technical field [0001] The invention relates to the field of biological products, in particular to an expressed protein for detecting bovine viral diarrhea virus antibody, an ELISA kit, a preparation method and application thereof. Background technique [0002] Bovine viral diarrhea virus (BVDV) is the main pathogen that causes bovine viral diarrhea disease (BVD). It mainly infects cattle, but also other organisms such as pigs, sheep, and camels. It has a wide range of hosts. Clinically, the main symptoms are diarrhea, growth and reproduction disorder, immunosuppression, acute or chronic mucosal disease and persistent infection. BVDV belongs to the genus Pestivirus of the family Flaviviridae, and has high homology with the other two swine fever viruses and border disease viruses of the Pestivirus genus, with structural and antigenic correlations, and serological Cross reaction. According to different typing criteria, BVDV can be divided into one serotype, two biotypes (cyt...

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
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/558
CPCG01N33/56983G01N33/558
Inventor 温永俊月英温永胜杨洋何芳
Owner 内蒙古元山生物科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com