pH35GC Vector
pH35GC Vector
Cat. No. FNK-IN3-VEC10
Store at -20°C
Size 10µg
Description
This is a binary vector series cloned with Gateway® system. In transformation of plants by the Agrobacterium method, it can be used for localization analysis in cells using fusion with a fluorescent protein, promoter analysis by expression of a fluorescent protein or GUS, etc. There is also a high expression construct that allows hygromycin resistance selection of plants.
It is a type of Gateway® binary vector that adds a fluorescent protein (CFP) to the C-terminal side and can be used for analysis of intracellular localization of target genes. You can use it at ease because it has been used at RIKEN Plant Science Research Center.
|
Name |
Position |
|
Left border (LB) |
1-26 |
|
NOS promoter (PNOS) |
118-301 |
|
HPT |
367-1392 |
|
RbcS terminator (RbcST) |
1411-2150 |
|
35S promoter (P35S) |
2187-3021 |
|
attR1 |
3049-3173 |
|
CmR |
3282-3941 |
|
ccdB |
4283-4588 |
|
attR2 |
4629-4753 |
|
Enhanced CFP (ECFP) |
4767-5486 |
|
NOS terminator (NOST) |
5508-5763 |
|
Right border (RB) |
6094-6118 |
Feature
- High expression by CaMV 35S promoter
- CFP can be linked to the C-terminal side of the Gateway® cassette to express the target gene as a CFP fusion protein and observe the intracellular localization
- Selection marker for E. coli / Agrobacterium: SpR (spectinomycin / streptomycin resistance)
- Selection marker of plant: HPT (hygromycin 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.
