pFAST-G02 Vector

Product#: IN3-VEC32
$2,692.00

Type:

  • Academic
  • Commercial
Availability:
Ships in 1-2 weeks

pFAST-G02 Vector(Academic / Commercial Entities)

Cat. No. IN3-VEC32
Store at -20°C
Size 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.

in3-vec32_fig1jpg.jpg 

It is a Gateway® binary vector for high gene expression, and can be used for functional analysis of genes and preparation of plants with high expression of target gene. 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)

117-373

BAR

393-944

NOS promoter (PNOS)

965-1237

OLE1 promoter (POLE1)

1369-2812

OLE1-GFP fusion protein (OLE1-GFP)

2813-4342

35S terminator (T35S)

4626-4445

35S promoter (P35S)

5245-6076

attR1

6175-6299

CmR

6408-7067

ccdB

7409-7714

 attR2

7755-7879

35S terminator (T35S)

7978-8097

Right border (RB)

8193-8217

Feature

  • High expression by CaMV 35S promoter
  • Gateway® cassette is linked between CaMV35S promoter and CaMV35S terminator
    • 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: Bar (Bialaphos resistant)

Form

10mM Tris-HCl (pH 7.4), 1mM EDTA

How to use
  1. 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)
  2. 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)
  3. Transform the expression clone prepared in 2. into Agrobacterium and infect the plant.
(For the selection of plants, please observe the GFP fluorescence of seeds or use a designated drug 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

Reference
  • Shimada T et al., Plant J. (2010) 61 (3): 519-528.
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