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

Soybean mosaic virus infectious clone vector capable of infecting tobacco, agrobacterium strain and application of vector and agrobacterium strain

A technology of soybean mosaic virus and cloning vector, applied in the field of genetic engineering, to achieve the effect of promoting the excavation process

Active Publication Date: 2020-07-24
NANJING AGRICULTURAL UNIVERSITY
View PDF3 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because few strains of soybean mosaic virus can infect tobacco, so in the research of soybean mosaic virus, there are few applications to tobacco

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
  • Soybean mosaic virus infectious clone vector capable of infecting tobacco, agrobacterium strain and application of vector and agrobacterium strain
  • Soybean mosaic virus infectious clone vector capable of infecting tobacco, agrobacterium strain and application of vector and agrobacterium strain
  • Soybean mosaic virus infectious clone vector capable of infecting tobacco, agrobacterium strain and application of vector and agrobacterium strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: Construction of soybean mosaic virus infective cloning vector pCB301-SMV (SC7)

[0030] 1. RNA extraction and reverse transcription of SMV (SC7) isolate

[0031] RNA was extracted using trizol reagent: take 1 leaf inoculated with SMV (SC7) and put it into a 2ml centrifuge tube, add liquid nitrogen, and use a grinding rod to quickly grind it into powder; add 1ml trizol to shake and mix, and let stand at room temperature for 5 minutes Add 200ml of chloroform and shake vigorously for 15sec, and let stand at room temperature for 2min; centrifuge at 12,000rcf for 10min in a 4°C centrifuge, absorb 500μl of the supernatant, add it to 500μl of isopropanol and mix well, let stand for 5min; centrifuge at 12,000rcf for 5min in a 4°C centrifuge, Discard the supernatant; add 1ml of 75% ethanol to vortex for 1min, centrifuge at 12000rcf for 1min at 4°C, discard the supernatant, and let stand at room temperature for 5min to fully evaporate the ethanol. Add 50 μl of wate...

Embodiment 2

[0036] Embodiment 2: the preparation of the bacterial strain ArgSMV (SC7) that Agrobacterium forms and the infection to Nicotiana benthamiana

[0037] 1. Agrobacterium freeze-thaw method to transform the invasive cloning vector pCB301-SMV(SC7)

[0038] Transform the invasive cloning vector pCB301-SMV(SC7) into the Agrobacterium strain EHA105 using the freeze-thaw method. The specific operation is as follows: take out 1 competent EHA105 and place it on ice to thaw, add 5 μl of pCB301-SMV(SC7) plasmid and Mix well, freeze in liquid nitrogen for 2 minutes, then place in a 37°C water bath for 3 minutes, add 800 μl of YEP medium to recover for 3 hours, take 100 μl and spread evenly on the YEB medium containing Kan, Str and Rif antibiotics, and incubate at 28°C for 3 sky. That is to make the Agrobacterium strain ArgSMV (SC7).

[0039] 2. Expansion culture of Agrobacterium bacteria liquid and preparation of injection

[0040] Pick a single colony on the above YEB plate, put it int...

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 soybean mosaic virus infectious clone vector capable of infecting tobacco, an agrobacterium strain and application of the s vector and the agrobacterium strain. The sequenceof the soybean mosaic virus infectious clone vector capable of infecting tobacco is shown as SEQ ID NO.1. The soybean mosaic virus infectious clone vector pCB301-SMV (SC7) is applied to tobacco infection. According to the invention, a soybean mosaic virus strain (SC7) capable of infecting tobacco is selected to construct an infectious clone vector, agrobacterium tumefaciens is transformed and injected into the tobacco, and then systematic infection is successfully formed on the tobacco. Therefore, a solid and reliable foundation is laid for the application of tobacco in the aspect of soybean mosaic virus research.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and relates to soybean mosaic virus infective cloning vectors capable of infecting tobacco, agrobacterium strains and applications thereof. Background technique [0002] Soybean mosaic virus (SMV) belongs to the genus Potatovirus, and its genome is a positive-sense RNA strand with a size of about 9.6 kb, including 5' and 3' non-coding regions and an intermediate coding sequence. A polypeptide translated from its coding sequence can form 10 mature viral proteins through self-cleavage. The host of soybean mosaic virus is relatively narrow, but its epidemic range is very wide, and it occurs to varying degrees in the main soybean producing areas of the world. After soybeans are infected with soybean mosaic virus, symptoms such as mosaic leaves, shrunken leaves, and dead tops will occur, resulting in hindered plant growth and development, and reduced yield and quality. [0003] Studying the pathog...

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): C12N15/82C12N15/40C12N1/21A01H5/00A01H6/82C12R1/01
CPCC12N15/8205C07K14/005C12N2770/34022
Inventor 智海剑殷金龙王丽群刘慧晋彤彤李凯
Owner NANJING AGRICULTURAL UNIVERSITY
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