関連する実験動画
Updated: Feb 13, 2026

08:38
Scalable Transfection of Maize Mesophyll Protoplasts
Published on: June 23, 2023
3.7K
トウモロコシメソフィルの細胞からの原生体の分離と感染
Lauren A Higa1, Taren Bouwman1, Zhi-Yan Du1
1Department of Molecular Biosciences & Bioengineering, University of Hawaii at Mānoa, Honolulu, HI, USA.
Bio-protocol
|February 12, 2026
まとめ
この研究は,トウモロコシの原生体分離とCRISPR/Cas9遺伝子編集のための最適化されたプロトコルを提示しています. この新しい方法は,プロトプラストの収穫量,生存能力,およびトランスフェクション効率を向上させ,植物研究を加速します.
科学分野:
- 植物分子生物学 植物分子生物学
- 農業バイオテクノロジー 農業バイオテクノロジー
背景:
- プロトプラストシステムは,植物研究と遺伝子編集の検証に不可欠です.
- 未熟の胚を介してトウモロコシの変容は遅いもので,資源が密集しています.
- 現存するトウモロコシ原型プラストプロトコルは,回復,変異,および生存能力において一貫性がない.
研究 の 目的:
- トウモロコシメソフィルの原生体分離とPEG媒介の転移のための最適化されたプロトコルを開発する.
- トウモロコシの遺伝子編集評価の効率,再現性,および速度を改善するために.
主な方法:
- 縦切りを用いてエチオレートされた苗からトウモロコシメソフィルの原生体分離を最適化しました.
- 還元されたプラズミドDNA (10μg) を使用したCRISPR/Cas9コンストラクタによるPEG4000媒介トランスフェクション.
- プロトプラストの生存期間を7日まで延長するために,洗浄と保存条件を修正しました.
主要な成果:
- 垂直切断によるプロトプラストの収量,一貫性,活性の改善.
- わずか10μgのプラズミドDNAで効率的なCRISPR/Cas9の配送が可能になりました.
- 改良された下流分析のために,変形した原形質の生存期間を7日まで延長しました.
- ターゲティングシーケンシングとCas-Analyzerを使用して,編集効率の定量化.
結論:
- このプロトコルは,トウモロコシの遺伝子編集を評価するための迅速で効率的で再現可能な方法を提供します.
- この方法論は,農作物の研究と植物分子生物学の研究を加速します.
関連する概念動画
Overview Of Cell Separation And Isolation
7.7K
Cell separation was first achieved in 1964 by S. H. Seal, who separated large tumor cells from the smaller blood cells using filtration. Two years later, Pohl and Hawk performed experiments on how cells respond differently to a nonuniform electric field based on the cell type. Such observations were the inception of cell separation methods, which allow isolating a single cell type from a heterogeneous sample.
7.7K
DNA Isolation
200.1K
DNA from cells is required for many biotechnology and research applications, such as molecular cloning. To remove and purify DNA from cells, researchers use various methods of DNA extraction. While the specifics of different protocols may vary, some general concepts underlie the process of DNA extraction.
200.1K
DNA Isolation
45.2K
DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
45.2K
C4 Pathway and CAM
49.3K
Most plants use the C3 pathway for carbon fixation. However, some plants, such as sugar cane, corn, and cacti that grow in hot conditions, use alternative pathways to fix carbon and conserve energy loss due to photorespiration. Photorespiration is the process that occurs when the oxygen concentration is high. Under such conditions, the rubisco enzyme in the Calvin cycle binds O2 instead of CO2, which halts photosynthesis and consumes energy.
C4 Pathway
The C4 pathway is used by plants such as...
C4 Pathway
The C4 pathway is used by plants such as...
49.3K
Anatomy of Chloroplasts
119.5K
Green algae and plants, including green stems and unripe fruit, harbor chloroplasts—the vital organelles where photosynthesis takes place. In plants, the highest density of chloroplasts is found in the mesophyll cells of leaves.
119.5K
Cyclic Processes And Isolated Systems
3.5K
A thermodynamic system with zero heat exchange and work is an isolated system. For these systems, the internal energy remains constant.
In the case of a non-isolated system, the change in the internal energy is zero only if the process is cyclic. A thermodynamic process is considered cyclic if the system undergoes a series of changes and returns to its initial state.
Consider a cyclic process that returns to its initial state, undergoing a four-step process. The heat transfer along each...
In the case of a non-isolated system, the change in the internal energy is zero only if the process is cyclic. A thermodynamic process is considered cyclic if the system undergoes a series of changes and returns to its initial state.
Consider a cyclic process that returns to its initial state, undergoing a four-step process. The heat transfer along each...
3.5K

