pFAST-G03 Vector (for Commercial Entities)
pFAST-G03 Vector(Commercail Entities / Academic)
Cat. No. IN3-VEC33
Store at -20°C
Size 10µg
| Product No. | Entity | Size | Link |
| FNK-IN3-VEC33-C | Commercial | 10µg |  |
| FNK-IN3-VEC33-A | Academic | 10µg | |
Description
Plasmid vector for plant transformation
This is a binary vector series cloned with Gateway® system. In transformation of Arabidopsis thaliana by the Agrobacterium method, it is possible to select transformed seeds under a fluorescent microscope by expressing seed-specific GFP, and the selection can be performed in a short time. It can be used for expression by any promoter, high expression, RNA silencing (RNAi) and promoter analysis.
This Gateway® binary vector for RNA silencing (RNAi) can be used for gene knockdown, etc. Seed-specific GFP fluorescence makes it possible to screen for transformation selection and homoheterogeneity. You can use it with confidence because it has been used at Kyoto University.
|
Name |
Position |
|
Left border (LB) |
1-26 |
|
NOS terminator (NOST) |
117-370 |
|
HPT |
568-1590 |
|
NOS promoter (PNOS) |
1605-1875 |
|
35S terminator (T35S) |
1950-2069 |
|
OLE1-GFP fusion protein (OLE1-GFP) |
2353-3882 |
|
OLE1 promoter (POLE1) |
3883-5326 |
|
35S terminator (T35S) 5365-5484 |
5365-5484 |
|
attR1 |
5583-5707 |
|
CmR |
5816-6475 |
|
ccdB |
6817-7122 |
|
attR2 |
7163-7287 |
|
Intron |
7355-7998 |
|
attR2 |
8013-8137 |
|
ccdB |
8178-8483 |
|
CmR |
8825-9484 |
|
attR1 |
9593-9717 |
|
35S promoter (P35S) |
9829-10657 |
|
Right border (RB) |
10863-10887 |
Feature
- A construct in which the inverted repeat, in which intron sequence is sandwiched by Gateway® cassette, is highly expressed by the CaMV 35 S promoter
- Can construct constructs that cause RNA silencing to the target sequence in one LR reaction
- In Arabidopsis thaliana, GFP is expressed in a seed-specific manner, and it is possible to select transformed seeds under a fluorescence microscope
- Can distinguish homoheteroes with fluorescence intensity
- Time to obtain T2 homozygotes is shortened compared to antibiotic selection
- Selection marker for E. coli / Agrobacterium: SpR (spectinomycin / streptomycin resistance)
- Selection marker of plant: HPT (hygromycin resistant)
Form
How to use
- Create an entry clone into which the target gene has been introduced * 1. There are several methods for producing entry clones, but we recommend BP reaction * 3 with PCR product and donor vector * 2. (Selection of E. coli is derived from resistant antibiotic of donor vector. In addition, competent cells should use DH5α * 4 with transformation efficiency of 10 ^ 10 or more)
- Perform LR * 5 reaction with the entry clone prepared in 1. and this vector (destination vector) to prepare an expression clone. (Please use the specified drug for each vector for selection in transformation, and use competent cells with DH5α * 4 with transformation efficiency of 10 ^ 10 or more)
- Transform the expression clone prepared in 2. into Agrobacterium and infect the plant.
* 1 Please refer to Thermo Fisher Scientific's HP for details.
* 2 Thermo Fisher Scientific pDONR Gateway / Zeo vector (Zeocin resistant)
* 3 Gateway BP Clonase Enzyme Mix manufactured by Thermo Fisher Scientific
* 4 Thermo Fisher Scientific Library Efficiency DH5α Competent Cell
* 5 Gateway LR Clonase enzyme mix manufactured by Thermo Fisher Scientific
Reference
- Shimada T et al., Plant J. (2010) 61 (3): 519-528.
