pBI-GW-NOS Vector

Product#: IN3-VEC1
$808.00
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pBI-GW-NOS Vector

Cat. No.
 IN3-VEC1 
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-vec1_fig1.jpg 

This promoterless Gateway® binary vector can be used to generate expression constructs with any promoter or to construct constructs for complementation tests by introducing genomic fragments. 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

attR1

4999-5123

CmR

5232-5891

ccdB

6233-6538

attR2

6579-6703

NOS terminator (NOST)

6822-7077

Left border (LB)

7716-7741

Feature

  • Link the promoterless Gateway® cassette upstream of the NOS terminator
  • 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. M Suzuki et al., J. Exp. Bot. (2009) 60 (7): 2055-2064.
  2. J Tang et al.,Plant J. (2010)61(3):456-466.
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