Ammopiptanthus mongolicus NAC1 gene promoter
A promoter and gene technology, applied in the field of Populus euphratica NAC1 gene promoter
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0024] Embodiment 1, Populus euphratica PeNAC1 gene promoter isolation
[0025] The promoter fragment of Populus euphratica PeNAC1 gene was isolated from the genomic DNA of Populus euphratica by PCR Walking method, and the specific operation method was carried out according to the instructions of the Genome Walking Kit method (TaKaRa, Japan).
[0026] The extraction of Populus euphratica DNA adopts CTAB method, and its steps are as follows:
[0027] (1) Add 800 μL of 2×CTAB extraction buffer [2g / 100ml CTAB, 1.4mol / L NaCl, 20mmol / L EDTA, 100mmol / L Tris·Cl (pH8.0)], 8μL mercaptoethanol in a 2mL centrifuge tube (1%, v / v), preheated at 65°C.
[0028] (2) Take about 0.15 g of leaves, grind them into frozen powder in liquid nitrogen.
[0029] (3) Transfer the frozen powder into a centrifuge tube, keep warm at 65°C for about 30 minutes, and shake gently twice during this time.
[0030] (4) Take out the centrifuge tube and cool to room temperature. Add an equal volume of chlorofor...
Embodiment 2
[0036] Example 2, Analysis of the promoter activity of Populus euphratica PeNAC1 gene
[0037] The promoter activity of the 1217bp 5' flanking sequence of Populus euphratica PeNAC1 gene was detected by transient expression experiment of onion epidermal cells.
[0038] Replace the 35S promoter in the plant expression vector pCAMBIA-1304 with the 1217bp 5' flanking sequence of the obtained Populus euphratica PeNAC1 gene, and insert it into the upstream of the reporter gene CUS and GFP to obtain a new plant expression vector pCAMBIA:PeNAC1P-GFP plasmid, which was transformed into In Agrobacterium LBA4404, the onion epidermal cells were infected by Agrobacterium, and the 1217bp 5'flanking sequence of Populus euphratica PeNAC1 gene obtained by transient expression experiment was detected whether it had promoter activity.
[0039] Using the enzyme cutting site analysis system (http: / / tools.neb.com / NEBcutter2 / index.php) on the NEB website to analyze all the enzyme cutting sites conta...
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