Mud crab bicistronic mRNA virus structural protein 1, monoclonal antibody thereof and application

A monoclonal antibody and structural protein technology, applied in antiviral immunoglobulin, applications, viral peptides, etc., can solve the problems that there are no related reports and no research on structural proteins

Inactive Publication Date: 2010-06-02
SUN YAT SEN UNIV
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Dicistrovirus (dicistrivirus) is a newly classified virus genus. With the deepening of marine virology, more and more dicistroviruses of aquatic organisms have been discovered, but their structural proteins have not been studied yet. There is no relevant report on the research on the structural protein of Scylla serrata dicistronic virus (MCDV)

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
  • Mud crab bicistronic mRNA virus structural protein 1, monoclonal antibody thereof and application
  • Mud crab bicistronic mRNA virus structural protein 1, monoclonal antibody thereof and application
  • Mud crab bicistronic mRNA virus structural protein 1, monoclonal antibody thereof and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The acquisition of embodiment 1 MCDV structural protein 1

[0036] In this example, the amino acid sequence and nucleotide sequence of structural protein 1 are obtained by purifying, N-terminal sequencing and analysis of MCDV, and biological analysis of structural protein 1 is carried out to predict its physical and chemical properties and conserved functional domains, which is the structure of MCDV To lay the foundation for protein function research, the specific steps are as follows.

[0037] 1. Purification of MCDV

[0038] (1) Weigh about 15 g of the gill sample of the diseased crab, and quickly grind it to powder under liquid nitrogen;

[0039] (2) Pour the ground powder into a 250ml centrifuge tube, add PBS in an amount 20 times the volume, and homogenate;

[0040] (3) Centrifuge the homogenized virus solution at 4°C to remove slag: centrifuge at 1000rpm for 20min; turn to centrifuge at 3000rpm for 3min; transfer the supernatant to a new tube, and after balancin...

Embodiment 2

[0052] Example 2 Sequence determination and biological analysis of MCDV structural protein 1

[0053] According to the SDS-PAGE electrophoresis results in Example 1, MCDV contains structural protein 1 (abbreviated as VP1) with a molecular weight of 53KD, structural protein 2 (abbreviated as VP2) with a molecular weight of 35KD and structural protein 3 (abbreviated as VP3) with a molecular weight of 22KD.

[0054] In this example, N-terminal sequencing and biological analysis of structural protein 1 were performed.

[0055] According to the routine operation of protein N-terminal amino acid sequencing in this field, first prepare the PVDF membrane of MCDV, select structural protein 1 on the PVDF membrane, and perform N-terminal amino acid sequencing.

[0056] Sequencing method: EDMAN degradation method.

[0057] Sequencing instrument: PROCISE491 sequencer from Applied Biosystems, USA.

[0058] Sequencing results:

[0059] The amino acid sequence of structural protein 1 is sh...

Embodiment 3

[0061] The preparation of the monoclonal antibody of embodiment 3MCDV structural protein 1

[0062] In this embodiment, the whole virus, that is, MCDV, is first used as an antigen to carry out an in vivo immune test, then HAT and HT are used to select hybridoma cells, and the positive monoclonal antibody is screened by indirect ELISA, and then the positive monoclonal antibody is screened by Western blot method. The monoclonal antibody of protein 1 was screened out, and finally 5 monoclonal antibodies capable of recognizing MCDV structural protein 1 were obtained, the specific steps are as follows.

[0063] 1. In vivo immunoassay

[0064] The antigen of in vivo immunization test adopts MCDV, and its operation adopts the routine operation when those skilled in the art carry out in vivo immunization test, carries out in vivo immunization test to BalB / C mouse and New Zealand white rabbit respectively, finally collects blood respectively (BalB / C mouse and New Zealand white rabbit)....

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
Molecular weightaaaaaaaaaa
Login to view more

Abstract

The invention discloses a mud crab bicistronic mRNA virus structural protein 1, a monoclonal antibody thereof and the application. The amino acid sequence of the mud crab bicistronic mRNA virus structural protein 1 is SEQ ID NO: 3 and the nucleotide sequence is SEQ ID NO: 1. By researching and analyzing MCDV, the invention firstly obtains the amino acid sequence, the nucleotide sequence and the monoclonal antibody of the structural protein 1, locates the structural protein 1 and provides a new research direction for researching virus of mud crab bicistronic mRNA. The monoclonal antibody of the structural protein 1 can be used in monitoring MCDV, preparing a virus kit for detecting MCDV and preparing a virus colloidal gold test paper for detecting MCDV.

Description

technical field [0001] The invention relates to the biological technology field of mud crab breeding, in particular to the bicistronic virus structural protein 1 of mud crab serrata and its monoclonal antibody and application. Background technique [0002] Scylla serrata (Scylla serrata, scientific name Mud crab), commonly known as blue crab, is mainly distributed in the tropical and subtropical waters of the Indo-Pacific Ocean. It is an edible crab with high economic value and is one of the famous and excellent breeding species. [0003] Viral diseases are the main pathogens in the breeding process of Scylla serrata, which cause serious economic losses in the breeding industry of Scylla serrata. Therefore, research on various viral diseases in the breeding process of Scylla serrata is an important topic at present. [0004] In 2004, two viruses were found in mud crabs suffering from "sleeping sickness" (SD) in Zhuhai: Mud crab dicistrivirus (MCDV) and reovirus (mudcrab reov...

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): C07K14/08C12N15/40C07K16/10G01N33/577C12R1/93
Inventor 张锐翁少萍何建国区宇洁董传甫
Owner SUN YAT SEN UNIV
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