Botanic vegetative growth maintenance gene system and establishment of vegetative growth maintenance plant and method for utilizing infinite growth capacity of seed plants
A vegetative growth and gene system technology, applied in botany equipment and methods, angiosperms/flowering plants, plant products, etc., can solve the problems of host cell death, expression, control mechanism not completely clear, cytotoxicity, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0098] Example 1 Construction of seed plant vegetative growth maintenance gene system (BTg-RBN)
[0099] (1) Cloning of Barstar gene:
[0100] Barstar gene was synthesized by PCR. The template is Bacillus amylobacter nuclear DNA, and the forward and reverse primers are: BTF1 5'- GTCCAATCTGCAGCCGTCCGAGACA -3', BTR1 5'- CGATTCAGGCGGCTG
[0101] ATATTCTCCA-3'.
[0102] The synthesized Barstar gene fragment was directly inserted into the pMD20-T vector of TaKaRa Company, confirmed by sequencing, and constructed into pMD20-BTg plasmid.
[0103] (2) Cloning of RBN promoter, coding sequence and terminator fragments:
[0104] The RBN gene is initially expressed in the Arabidopsis reproductive meristem AtREM1Gene promoter, Barnase gene coding sequence, 3A terminator consists of 3 fragments. These three gene fragments are all cloned by PCR technology.
[0105] Gene promoter: The source of the fragment sequence is AL161579 (82727-83643bp), with a length of 919bp; the restriction si...
Embodiment 2
[0115] Embodiment 2 Construction of plant expression vector of seed plant vegetative growth maintenance gene system (BTg-RBN)
[0116] Only when the constructed vegetative growth-maintaining gene is loaded into a plant expression vector and transferred into a plant, can it present a function in the plant and play a role.
[0117] A variety of plant control expression vectors have been established so far, each with its own characteristics, such as pBI121, pCAMBIA series and so on. In this embodiment, the preferred expression vector is pCAMBIA3301. The vector carries the left and right borders LB and RB of the T-DNA of the Ti plasmid of Agrobacterium tumefaciens, and can be transformed into plants by the method mediated by Agrobacterium tumefaciens; the selectable marker in the plants is Bar.
[0118] The establishment of the vegetative growth maintenance gene system RBN.BTg expression vector was completed by cutting out the RBN-BTg fragment from the cloning vector with HindⅢ a...
Embodiment 3
[0119] Example 3 Identification of vegetative growth-maintaining plant bodies
[0120] Confirmed by detecting the presence of the (AtREM1-Barnase-3A) gene fragment, or related fragments, in the nuclear genome of transformed plants. The identification procedure is the same as that of general transgenic plants, and the upstream and downstream primers used can be respectively: RB t F: 5'-TTAGTAGGATTCTGGTGTGTGGG-3'; RB t R: 5'-TTCGATTTTTAATAGATTCGGTTTT-3'. The fragment length is 611bp (including AtREM1 Part of the promoter and Barnase coding sequence and part of the terminator sequence of 3A).
[0121] For plants whose reproductive organs appear after many years of planting, the identified transgenic plant materials can be directly grafted on adult trees (with a control) to finally determine whether reproductive organs, growth characteristics and other morphological characteristics can be produced. Generally, it takes 2 to 3 years to identify whether the transformed plant is a...
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