pH35GC Vector

Product#: IN3-VEC10
$1,154.00
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pH35GC Vector

Cat. No
. IN3-VEC10
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.

in3-vec10 _fig1.jpg 

It is a type of Gateway® binary vector that adds a fluorescent protein (CFP) to the C-terminal side and can be used for analysis of intracellular localization of target genes. 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

HPT

367-1392

RbcS terminator (RbcST)

1411-2150

35S promoter (P35S)

2187-3021

attR1

3049-3173

CmR

3282-3941

ccdB

4283-4588

attR2

4629-4753

Enhanced CFP (ECFP)

4767-5486

NOS terminator (NOST)

5508-5763

Right border (RB)

6094-6118

Feature

  • High expression by CaMV 35S promoter
  • CFP can be linked to the C-terminal side of the Gateway® cassette to express the target gene as a CFP fusion protein and observe the intracellular localization
  • Selection marker for E. coli / Agrobacterium: SpR (spectinomycin / streptomycin resistance)
  • Selection marker of plant: HPT (hygromycin 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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