pBIDAVL-GWR1 Vector

pBIDAVL-GWR1 Vector

Cat. No. FNK-IN3-VEC7 
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.

A Gateway® binary vector for high expression in the sense direction, to which a specific sequence has been added for RT-PCR, and can be used for functional analysis of genes and production of plants with high expression of target genes. You can use it at ease because it has been used at RIKEN Plant Science Research Center.

Feature

• High expression by CaMV 35S promoter
• Link Gateway® cassette between 35S promoter and NOS terminator
  • By RT-PCR using a specific sequence (DAVL) inserted inside the polyA addition site, it is possible to confirm whether or not the RNA derived from the introduced gene is accumulated in the cell
• Use in combination with pBIDAVL-GWR2 allows construction of a sense orientation and antisense orientation from the same entry clone at one time
• 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.