pFAST-G04 Vector

Product#: IN3-VEC34
$2,692.00

Type:

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

pFAST-G04 Vector (Commercial Entities / Academic)

Cat. No. in3-vec34
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.

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A Gateway® binary vector that expresses a GFP-GUS fusion protein and can be used for promoter analysis, 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

NPTII

410-1204

NOS promoter (PNOS)

1217-1489

35S terminator (T35S)

1615-1734

OLE1-GFP fusion protein (OLE1-GFP)

2018-3547

OLE1 promoter (POLE1)

3548-4970

attR1

4980-5104

CmR

5213-5872

ccdB

6214-6519

attR2

6560-6684

GFP-GUS fusion protein (GFP-GUS)

6697-9228

35S terminator (T35S)

9338-9457

Right border (RB)

9553-9577

Feature

  • Link Gateway® cassette upstream of GFP-GUS fusion protein
  • Promoter analysis is possible by incorporating the target promoter into the Gateway® cassette
    • 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: NPTII (Kanamycin 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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