pBI-GW-NOS(GFP) Vector
pBI-GW-NOS(GFP) Vector
Cat. No. FNK-IN3-VEC4
Store at -20°C
Size 10µg
Description
This binary vector series is based on pBI vector and cloned using Gateway® system. Agrobacterium-mediated plant transformation can be used for expression with any promoter, high expression, and RNA silencing (RNAi). There is also a type that expresses GFP as a reporter or a type that adds a specific sequence for confirmation of RT-PCR.
Promoterless Gateway® binary vector with GFP as a reporter, it can be used to construct expression constructs with any promoter or construct for complementation tests by introducing genomic fragments. You can use it at ease because it has been used at RIKEN Plant Science Research Center.
|
Name |
Position |
|
Right border (RB) |
2454-2478 |
|
NOS promoter (PNOS) |
2519-2825 |
|
NPTII |
2838-3632 |
|
NOS terminator (NOST) |
4022-4277 |
|
35S promoter (P35S) |
4968-5334 |
|
GFP |
5429-6148 |
|
NOS terminator (NOST) |
6167-6419 |
|
attR1 |
6475-6599 |
|
CmR |
6708-7367 |
|
ccdB |
7709-8014 |
|
attR2 |
8055-8179 |
|
NOS terminator (NOST) |
8301-8556 |
|
Left border (LB) |
9195-9220 |
Feature
- Link the promoterless Gateway® cassette upstream of the NOS terminator
- Confirmation of gene transfer by sGFP (S65T) fluorescence is possible
- Selection marker of E. coli / Agrobacterium: NPTIII (Kanamycin resistance)
- Selection marker of plant: NPTII (Kanamycin resistant)
Form
10mM Tris-HCl (pH 7.4), 1mM EDTA
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.
(For plant selection, please use the drug specified for each vector)
* 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
Attention point
When creating a construct in which the target gene and the fluorescent protein are fused, design so that the codon frames match.
