In bread wheat comprehensive genome analysis is complicated by several factors such as a very large genome (∼17 Gbp), its polyploid nature and a high repetitive sequence content. Using flow cytometry allows to dissect the wheat giant genome to defined parts mainly its chromosomes and chromosome-arms and can help to solve this challenge by enabling the chromosome-based strategy. Flowsorted chromosomes were used already to construct several grain BAC libraries including libraries from chromosome 3B and chromosome arms IBS and 1RS. They became valuable resources for wheat physical mapping and positional cloning, and they had an insert size of 75-85 kb, as average. This insert size was a consequence of a limited amount of DNA obtained after a time-consuming sorting which enabled only one size-selection step. In the present work we compare the construction of the 3AS BAC library in respect to our new improved protocol which made the second size-selection step feasible and which was applied for the construction of a BAC library specific for chromosome arm 3DS, which showed an average insert size reaching 110 kb. A minimal tiling path of chromosome 3B has already been established. Construction of the 3AS- and 3DS-specific BAC libraries represent significant step toward completing BAC resources for the homoeologues chromosome group 3 of wheat and accelerate the development of sequence-ready physical contig maps and gene cloning, as well as comparative analyses aiming at revealing the genome changes accompanying the evolution of homoeologous chromosome group 3.
|Pages (from-to)||151 - 160|
|Number of pages||10|
|Journal||Journal of Genetics and Breeding|
|Publication status||Published - Jan 2007|
All Science Journal Classification (ASJC) codes
- Plant Science
Šafář, J., Šimková, H., Kubaláková, M., Suchánková, P., Číhalíková, J., Bartoš, J., ... Lucretti, S. (2007). Generating resources for genomics of wheat homoeologous chromosome group 3: 3AS- and 3DS-specific BAC libraries. Journal of Genetics and Breeding, 61(1-2), 151 - 160.