Reverse cross-multistage nerve tracing method
A nerve and tracer technology, applied in the field of virus neurotracer research, can solve problems such as weak PRV labeling efficiency and labeling effect, inability to infect primates, biosafety issues, etc., to achieve labeling efficiency and excellent effect, The effect of convenient subsequent transformation and short length
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 2
[0058] Example 2 Transgenic mice construction
[0059] C57BL / 6 mice as experimental animals.
[0060] The target carrier is primarily subjected to EMBRYONIC STEM Cell, embryonic stem cells, and ES cells below. Screening the correct recombinant ES cell clone (ie, the Successful ES cell clone).
[0061] Screening Policy: Select the Southern Blot method while using 5'Probe (probe), 3'Probe and Neoprobe for ES cell screening, 5 'end SoutherN enzyme cutting between 5' end LOXP sites and 3 × STOP elements Point, there is a 3 'side Southern enzyme sink downstream of Neo Cassette. If the correct recombination occurs, there will be two strips of wild-type and mutant types; if it is not correct, there will only be wild-type strips.
[0062] Restricted endonuclease Probe Wild-type (WT) amplification results Targeted amplification results HindIII 5’ 4.4KB 5.5KB EcorV 3’ 11.5kb 9.5kb XbaI 3'neo - 5.7kb
[0063] When the NEO gene is integrated into eukaryo...
Embodiment 3
[0082] Example 3 Transgenic animals and G protein lack of rabies virus binding to neural trace
[0083] A gt mouse which was capable of expressing a G protein as in Example 2 was an infected animal, and the GT rats were inoculated (drip nose or muscle injection) using a G protein defective CVS-N2C strain RV.
[0084] Such as Figure 9 As shown, 1 is the original genotype of the strain, carrying the five viral proteins of N, P, M, G, and L. 2 is a genotype after the lack of G protein of the strain, and the G protein is replaced by GFP (green fluorescent protein).
[0085] After infection, any one of the cells infected by the gt mouse is a G protein defective CVS-N2C strain RV, which restores the infection of street toxic RV, and simulates the natural infection process of RV downtown, you can pass G. The fluorescent gene expression in the protein defective strain itself observes the infection and dissemination of neurons of transgenic animals that provide G protein.
Embodiment 4
[0086] Example 4 Nerve tracer using a G protein defective RV
[0087] Using SAD-B19- Δg-GFP RV of the G-protein defective (ΔG) as a tracery virus tool, injecting the strand of the strand of the gt mouse described in Example 2, the viral titer is 2 * 10 ^ 8pfu / ml, the injection volume is 1 μl.
[0088] After 24 days, the gt mouse was perfused, and the polymethylene formaldehyde was fixed to take the brain tissue, dehydrated, and finally the topic slices of the brain sample were treated. It was observed whether the brain was observed by the G protein lack of RV trails. Virus infection and tag.
[0089] Because the virus tool genome carries the expression gene GFP of the green fluorescent protein, the infected neurons will be labeled with green fluorescent. According to the experimental results, we have found that the G protein-deficient RV trace virus tool that cannot be spread and infected in wild murine can be infected with muscle infection and spread back to the brain, infected...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
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