pBGCN Vector

Product#: IN3-VEC15
$1,731.00
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pBGCN Vector

Cat. No.
 IN3-VEC15
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.

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A promoterless Gateway® binary vector in which a fluorescent protein (CFP) is localized in the nucleus and can be used for promoter analysis. 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

BAR

351-902

RbcS terminator (RbcST)

1035-1774

attR1

1801-1925

CmR

2034-2693

ccdB

3035-3340

attR2

3381-3505

Nuclear-localized ECFP (ECFP-nuc)

3510-4340

NOS terminator (NOST)

4357-4612

Right border (RB)

4943-4967

Feature

  • Connect Gateway® cassette upstream of CFP with nuclear localization signal
  • Promoter analysis is possible by incorporating the target promoter into the Gateway® cassette
  • 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 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.

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