pBI-sense,anti sense GW(GFP) Vector

Product#: IN3-VEC6
$1,154.00
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pBI-sense,anti sense GW(GFP) Vector

Cat. No. IN3-VEC6
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-vec6_fig1.jpg 

A Gateway® binary vector for RNA silencing (RNAi) that has GFP as a reporter and 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

35S promoter (P35S)

4968-5334

GFP

5429-6148

NOS terminator (NOST)

6167-6419

35SS promoter (P35SS)

6448-7141

attR1

7173-7297

CmR

7406-8065

ccdB

8407-8712

attR2

8753-8877

Intron

8905-9193

attR2

9220-9344

ccdB

9385-9690

CmR

10032-10691

attR1

10800-10924

NOS terminator (NOST)

10992-11247

Left border (LB)

11886-11911

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
  • 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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