Method for structuring Chinese wheat mosaic virus infectivity clone and application of method

A technology of wheat mosaic virus and virus genome, applied in biochemical equipment and methods, using vectors to introduce foreign genetic material, recombinant DNA technology, etc., can solve the problems of hindering replication, understanding of pathogenic mechanism of assembly, and limiting identification of wheat resistance, etc. problem, to achieve the effect that is beneficial to wheat resistance and screening

Pending Publication Date: 2016-05-04
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
View PDF1 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The full-length cDNA cloning of the virus is an important technical means for in-depth understanding of the biological characteristics of the virus. The lack of CWMV invasive clones not only seriously hinders people's understanding of the mechanisms of CWMV infection, rep...

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
  • Method for structuring Chinese wheat mosaic virus infectivity clone and application of method
  • Method for structuring Chinese wheat mosaic virus infectivity clone and application of method
  • Method for structuring Chinese wheat mosaic virus infectivity clone and application of method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0021] Embodiment: A simple and fast method for constructing Chinese wheat mosaic virus invasive clone and its infection activity analysis to wheat, specific steps:

[0022] Primer design: A total of 4 primers were designed based on the RNA1 and RNA2 sequences of the Chinese wheat mosaic virus genome fragments determined by our laboratory. for amplifying RNA2;

[0023] Total RNA extraction of virus genome: take the extract containing 50-100 μl of CWMV particles, use Trizol reagent (Invitrogen) to extract total RNA of virus genome according to the reagent instructions, transfer the virus extract into a 1.5ml new centrifuge tube, add 1ml Trizol reagent (Invitrogen), After shaking and mixing, add 0.2ml chloroform, let stand for 3-5min after mixing, centrifuge at 12000g for 10min, take the supernatant to a new centrifuge tube and add an equal volume of isopropanol, mix and centrifuge at 12000g for 10min at 4°C, discard The supernatant and the pellet were washed twice with 70% eth...

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 relates to a method for easily and quickly structuring a Chinese wheat mosaic virus (CWMV) whole-length cDNA infectivity clone. One-step method RT-PCR amplification is carried out with a purified viral genome RNA as a template by applying one set of viral genome specific primers, wherein the number of the primers is four, and the primers are P1F, P1R, P2F and P2R respectively; an amplification product is purified and then connected to a carrier. It is shown through biological activity analysis that the CWMV whole-length cDNA clone obtained through the method can successfully infect wheat and other plants and can be replicated, subjected to system movement and assembled in plant bodies, and thus it is indicated that the clone has the infection activity and can be used for research of related virus function genomes and analysis of plant disease resistance.

Description

technical field [0001] The invention relates to a method for constructing a full-length cDNA infectious clone of an important crop virus and its application field, in particular to a method for constructing an infectious clone of Chinese wheat mosaic virus and its application. Background technique [0002] Soil-borne viral diseases of cereal crops occur widely in the world and have been difficult to control. At the end of the last century, our laboratory first discovered a soil-borne virus that caused wheat mosaic virus in Shandong, my country. Molecular identification showed that the soil-borne virus was a new species of plant virus, named Chinese wheat mosaic virus (Chinese wheat mosaic virus, CWMV). The new species of the virus has been classified into the genus Furovirus of the family Virgaviridae. CWMV can be transmitted by the obligate root-parasitic Polymyxagraminis and can survive for several years in the dormant spores of the fungal vector. Therefore, it is difficu...

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): C12N15/82C12N15/66
CPCC12N15/8203C12N15/66
Inventor 张恒木羊健张芬李静陈剑平
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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