Technology and Research Platforms

Crop Transformation (BRACT)

Our Crop Transformation Platform offers transformation and genome editing in a range of species including wheat, barley and Brassica crops.

We are unique in that we can offer a complete resource from experimental design and construct assembly through to transformation and screening of the plants developed. We can also provide training, ready prepared standard constructs, and help with grant proposals.

Our platform can help to advance research in many areas of plant science by providing functional characterisation of genes of interest and by providing knock-out mutants using CRISPR/Cas9 based technologies.

We offer;

  • Crop Genome editing
  • Wheat transformation
  • Barley transformation
  • Brassica transformation
  • Transformation constructs
  • Genome editing constructs
  • Plant transformation resources
  • Plant genome editing resources
  • CRISPR/Cas9 resources

We focus on providing resources to the academic community but may be able to help with some commercial projects too.

Each project is unique and is costed individually but as a guide;

  • Wheat transformation = £5,807/construct
  • Barley transformation = £3,056/construct
  • Brassica transformation = £3,875/construct
  • Constructs can be designed and assembled for £2,651 for standard constructs and £3,672 for CRISPR constructs

As projects run over several months contact us as early as possible to discuss potential work.

Protocols

Simple and efficient protocols developed by BRACT for routine Agrobacterium tumefaciens mediated transformation with recommended genotypes are detailed below for each of the crop species.

BRACT has a 100% success rate in technology transfer of these protocols to other labs.

Contact us for further information regarding protocols or for the opportunity to come and train at our facility.

Barley

Brassica

Wheat

Constructs

A series of tried and tested pBRACT vectors have been designed and tailored for each of the crop species.

These are available to researchers under MTA. Prices are available on request.

The pBRACT vectors are based on pGreen, which is a small, versatile vector designed for easy manipulation in E.coli with a high copy number.

To enable the small size of pGreen, the pSa origin of replication required for replication in Agrobacterium, is separated into its’ two distinct functions. The replication origin (ori) is present on pGreen, and the trans-acting replicase gene (RepA) is present on an additional vector, named pSoup. Both vectors are required in Agrobacterium for pGreen (or pBRACT) to replicate.

The pBRACT vectors have been designed as Gateway™ destination vectors. They therefore use Gateway™ cloning as the method for gene introduction, which is simple and reliable.

To use the Gateway ™ recombination reactions your gene of interest first needs to be cloned into a pENTRY vector. Your gene can then be introduced into any one of the pBRACT destination vectors using a one tube reaction.

Cereal constructs (suitable for Barley and Wheat)

Name

Function

Description

Word file

LB

RB
pBract202 – 35S Hygint – pBract202seq&map
pBract203 – 35S Hygint Gatecass pBract203seq&map
pBract208 – 35S Hygint Gatecass pBract208seq&map
pBract209 – 35S Hygint Gatecass-r pBract209seq&map
pBract204 control 35S Hygint Ubi GUSint pBract204seq&map
pBract207 RNAi silencing 35S Hygint Ubi RNAi pBract207seq&map
pBract210 control 35S Hygint Ubi LUCint pBract210seq&map
pBract211 over-expression without Gateway 35S Hygint Ubi-nos pBract211seq&map
pBract214 over-expression with Gateway 35S Hygint Ubi-nos pBract214-ss
pBract001 clean gene – Ubi GUSint pBract001seq&map
pSoup20B clean gene 35S Hygint – pSoup20Bseq&map

Brassica constructs

Name

Function

Description

Word file

35S Kan

35S Gatecass
pBract102 – 35S Kan – pBract102seq&map
pBract103 – 35S Kan Gatecass pBract103seq&map
pBract507 RNAi silencing nos Kan 35S RNAi pBract507seq&map
pBract114 over-expression
pBract104 GUS reporter 35S Kan 35S
Gus-Int
A new series of Bract constructs will be available here shortly.

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