pBI-GW-NOS(GFP) Vector

Product#: IN3-VEC4
$1,154.00
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pBI-GW-NOS(GFP) Vector

Cat. No.
 IN3-VEC4
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-vec4_fig1.jpg 

Promoterless Gateway® binary vector with GFP as a reporter, it can be used to construct expression constructs with any promoter or construct 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

35S promoter (P35S)

4968-5334

GFP

5429-6148

NOS terminator (NOST)

6167-6419

attR1

6475-6599

CmR

6708-7367

ccdB

7709-8014

attR2

8055-8179

NOS terminator (NOST)

8301-8556

Left border (LB)

9195-9220

Feature

  • Link the promoterless Gateway® cassette upstream of the NOS terminator
  • Confirmation of gene transfer by sGFP (S65T) fluorescence is possible
  • 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.

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