Description
EHA105 (pSuperAgro® v4tet) Agrobacterium Electrocompetent Cells
GoldBio's EHA105 (pSuperAgro® v4tet) Electrocompetent Agrobacterium cells combine the highly virulent EHA105 strain with the enhanced pSuperAgro® v4tet helper plasmid to maximize T-DNA delivery and improve plant transformation workflows. Optimized for electroporation, these transformation-ready cells provide researchers with a reliable platform for introducing binary vectors into Agrobacterium tumefaciens prior to Agrobacterium-mediated gene transfer.
EHA105 is a disarmed derivative of the hypervirulent A281 strain and is widely used for stable transformation of numerous monocot and dicot plant species, including Arabidopsis, tobacco, tomato, soybean, rice, maize, and many additional crops. Its high virulence and broad host range make it one of the most commonly selected strains for plant biotechnology, functional genomics, CRISPR/Cas genome editing, and transgenic crop development.
The pSuperAgro® v4tet helper plasmid enhances Agrobacterium-mediated transformation by simultaneously expressing ACC deaminase (AcdS) and GABA transaminase (GabT) from a single lac promoter. These enzymes reduce ethylene production and metabolize γ-aminobutyric acid (GABA), two important components of the plant defense response that can limit T-DNA transfer during co-cultivation. By reducing these inhibitory responses, pSuperAgro® v4tet enhances Agrobacterium-mediated T-DNA transfer and has been shown to improve both transient and stable transformation performance across multiple plant species.
The v4tet helper plasmid utilizes a tetracycline-selectable marker for convenient plasmid maintenance while preserving compatibility with common binary vector selection systems. Combined with the proven EHA105 background, this creates a robust platform for high-efficiency plant transformation.
Each kit includes high-efficiency electrocompetent EHA105 (pSuperAgro® v4tet) cells, Agrobacterium Recovery Medium, and pCAMBIA1391z control DNA for validating transformation performance.
Applications
- Agrobacterium-mediated plant transformation
- Stable and transient gene expression
- CRISPR/Cas genome editing
- Binary vector introduction
- Plant synthetic biology
- Functional genomics
- Gene overexpression and RNAi studies
- Transformation of monocot and dicot plant species
Key Features
- High-efficiency EHA105 electrocompetent Agrobacterium tumefaciens
- Enhanced Agrobacterium-mediated T-DNA transfer using pSuperAgro® v4tet
- Dual AcdS and GabT activities reduce plant defense responses
- Tetracycline-selectable pSuperAgro® v4tet helper plasmid
- Suitable for transformation of a broad range of plant species
- Animal-origin-free manufacturing
- Includes Agrobacterium Recovery Medium and pCAMBIA1391z control DNA
These products are sold under license by GoldBio and require a signed agreement before fulfilment. The purchase of this product includes a 1-year subscription to use pSuperAgro® in your research.
GoldBio’s EHA105 Agrobacterium strain was generated, and primary clone supplied by Dr. Elizabeth Hood.
Kit Components
- Competent Cells
- 1 x 12 mL Recovery Media
- 1 x 10 µL Control Plasmid (pCAMBIA1391z Control, 500 pg/µL)
Reagents Needed for One Reaction
- EHA105 (pSuperAgro® v4tet) Electrocompetent Agrobacterium: 25 µL
- DNA (pCAMBIA1391z Control, 500 pg/µL): 1 µL
- Recovery medium: 1 mL
Storage/Handling
This product may be shipped on dry ice. EHA105 Agrobacterium Electrocompetent cells should be stored at -80°C, pCAMBIA1391z Control DNA should be stored at -20°C and recovery medium should be stored at 4°C immediately upon arrival. When stored under the recommended conditions and handled correctly, these products should be stable for at least 1 year from the date of receipt.
Quality Control
Transformation efficiency is tested by using the pCAMBIA1391z control DNA supplied with the kit and using the protocol given below. Transformation efficiency should be ≥1 x 10 7 CFU/µg pCAMBIA1391z DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.
General Guidelines
- Handle competent cells gently as they are highly sensitive to changes in temperature or mechanical lysis caused by pipetting.
- Thaw competent cells on ice and transform cells immediately following thawing. After adding DNA, mix by tapping the tube gently. Do not mix cells by pipetting or vortexing.
Note: A high-voltage electroporation apparatus capable of generating field strengths of 16 kV/cm is required.
Calculation of Transformation Efficiency
Transformation Efficiency (TE) is defined as the number of colony forming units (cfu) produced by transforming 1 µg of plasmid into a given volume of competent cells.
- TE = Colonies/µg/Dilution
- Colonies = the number of colonies counted
- µg = amount of DNA transformed in µg
- Dilution = total dilution of the DNA before plating
Example: Transform 1 µl of (10 pg/µl) control plasmid into 25 µl of cells, add 975 µl of Recovery Medium. Dilute 10 µl of this in 990 µl of Recovery Medium and plate 50 µl. Count the colonies on the plate the next day. If you count 250 colonies, the TE is calculated as follows:
Colonies = 250
µg of DNA = 0.00001
Dilution = 10/1000 x 50/1000 = 0.0005
TE = 250/0.00001/0.0005 = 5.0 × 1010
pSuperAgro® is a registered trademark of Gold Biotechnology, Inc. Registration Number: 7,996,391
