UCE: A uracil excision (USER)-based toolbox for transformation of cereals.

ABSTRACT:

Background

Cloning of gene casettes and other DNA sequences into the conventional vectors for biolistic or Agrobacterium-mediated transformation is hampered by a limited amount of unique restriction sites and by the difficulties often encountered when ligating small single strand DNA overhangs. These problems are obviated by "The Uracil Specific Excision Reagent (USER)" technology (New England Biolabs) which thus offers a new and very time-efficient method for engineering of big and complex plasmids.

Results

By application of the USER system, we engineered a collection of binary vectors, termed UCE (USER cereal), ready for use in cloning of complex constructs into the T-DNA. A series of the vectors were tested and shown to perform successfully in Agrobacterium-mediated trans

SUBMITTER: Hebelstrup KH 

PROVIDER: S-EPMC2892451 | biostudies-literature | 2010 Jun

REPOSITORIES: biostudies-literature