Genomic Services

Crop Transformation (BRACT)

BRACT (Biotechnology Resources for Arable Crop Transformation) is a research facility based at the John Innes Centre providing the wider research community with access to a range of crop transformation and genome editing resources, such as;

  • Development of new targeted gene knockout resources: CRISPR/Cas9 technology
  • Training facilities: hosting visitors for hands-on training at the John Innes Centre
  • Constructs: developed specifically for Brassica and the Cereals – the pBRACT series of vectors

The facility operates on a ‘not-for-profit’, cost recovery basis and supports researchers worldwide.

BRACT services at the John Innes Centre, include;

  • Wheat transformation
  • Barley transformation
  • Brassica oleracea transformation
  • Brassica napus transformation
  • Medicago transformation
  • RNA-guided Cas9 mediated genome editing
  • Constructs including design and assembly
  • Training courses
  • Opportunities to develop collaborative projects
  • Transformation protocols

“The BRACT platform has a long history of providing efficient crop transformation to the research community. Recently we have demonstrated efficient genome editing in barley and Brassica crops and we are now able to provide targeted gene knockouts using RNA-guided Cas9 in addition to other transformation resources”

– Wendy Harwood, Head of Crop Transformation

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. Prices are available on request.

Barley

Brassica

Wheat

A new wheat transformation protocol will be available here shortly.

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

Brassica constructs

Name

Function

Description

Word file

35S Kan

35S Gatecass

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