Agrobacterium-mediated pythium oligadrum gene transformation method
A genetic transformation method and technology mediated by Agrobacterium, applied in the field of genetic transformation of Pythium oligandrum mediated by Agrobacterium, can solve the problems of reducing the transformation rate, complicated process, inability to transform, etc., to restore growth vitality and facilitate picking , The effect of simplifying the operation process
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0033] Example 1 Agrobacterium-mediated genetic transformation experiment using Pythium oligandrum mycelial spore suspension
[0034] (1) Inoculate Pythium oligandrum hyphae on PDA plate medium, culture at 26°C for 6 days, wash the plate with 2-4mL induction medium IM, wash off the Pythium oligandrum spores and part of the mycelium on the plate, Make a concentration of 1×10 7 spore mycelia suspension per mL.
[0035] (2) Pick a single clone from the Agrobacterium engineering strain resistance plate containing the target gene carrier, and put it in 5ml LB (50mg / L rifampicin Rif + 50mg / L kanamycin Kan) liquid medium at 28°C , shake culture at 200rmp for 20h; pipette 2mL of the above bacterial solution into 20mL LB (50mg / L Rif + 50mg / L Kan + 300uM acetosyringone) liquid medium and culture at 28℃, 240rmp until OD 600 2.5; 4°C, centrifuge at 5000rmp for 10min, collect the bacteria, resuspend and dilute the bacteria with IM osmosis medium (PDB+300 uM acetosyringone, pH 5.0) to mak...
Embodiment 2
[0042]Example 2 Experiment on the influence of different co-cultivation time on genetic transformation.
[0043] With reference to the method described in Example 1, Agrobacterium bacterium liquid and Pythium oligandrum mycelial spore suspension are co-cultivated for different times, and the optimal co-cultivation time is determined according to the PCR detection results, and the experimental results are shown in the table below:
[0044]
[0045] It can be seen from the experimental results that a good transformation effect can be obtained when the Agrobacterium bacterial liquid is co-cultivated with Pythium oligandrum mycelia and spore suspension for 48-96 hours.
Embodiment 3
[0046] Example 3 Experiment on the influence of recovery culture on genetic transformation.
[0047] With reference to the method described in Example 1, whether Agrobacterium bacterium liquid and Pythium oligandrum hyphae and spore suspension are co-cultivated and whether they experience recovery and recovery time, the experimental results are shown in the table below:
[0048]
[0049] It can be seen from the experimental results that the recovery culture stage can well promote the recovery of growth vigor of the infected Pythium oligandrum and increase the transformation effect, and the recovery culture period can be 24-48 hours.
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