pFAST-G03 Vector

Product#: IN3-VEC33
$2,423.00

Type:

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

pFAST-G03 Vector(Commercail Entities / Academic)

Cat. No. IN3-VEC33
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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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

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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