pBI sense, antisense GW Vector

Product#: IN3-VEC3
$808.00
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pBI sense,antisense GW Vector

Cat. No.
 IN3-VEC3 
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.

in3-vec3_fig1.jpg 

This Gateway® binary vector for RNA silencing (RNAi) can be used for gene knockdown, etc. 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

35SS promoter (P35SS)

4973-5666

attR1

5698-5822

CmR

5931-6590

ccdB

6932-7237

attR2

7278-7402

Intron

7430-7718

attR2

7745-7869

ccdB

8091-8396

CmR

9092-9751

attR1

9325-9449

NOS terminator (NOST)

9517-9772

Left border (LB)

10411-10436

Feature

  • A construct in which the inverted repeat in which the intron sequence is sandwiched by the Gateway® cassette is highly expressed by the CaMV 35 S promoter (35 SS promoter) in which the enhancer region is duplicated.
  • Can construct constructs that cause RNA silencing to the target sequence in one LR reaction
  • 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

  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 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.

Reference 
(used as pBCR112 in the reference)

  1. K Nemoto et al., FEBS Let. (2009) 583 (2): 487-492
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