BAC Libraries from Wheat Chromosome 7D: Efficient Tool for Positional Cloning of Aphid Resistance Genes

0364122 - ÚEB 2012 RIV US eng J - Journal Article
Šimková, Hana - Šafář, Jan - Kubaláková, Marie - Suchánková, Pavla - Čihalíková, Jarmila - Robert-Quatre, Heda - Azhaguvel, P. - Weng, Y. Q. - Peng, J. - Lapitan, N. L. V. - Ma, Y. Q. - You, F. M. - Luo, M. C. - Bartoš, Jan - Doležel, Jaroslav
BAC Libraries from Wheat Chromosome 7D: Efficient Tool for Positional Cloning of Aphid Resistance Genes.
Journal of Biomedicine and Biotechnology. -, č. 302543 (2011), s. 1-11. ISSN 1110-7243
R&D Projects: GA ČR GA521/07/1573; GA MŠMT(CZ) LC06004
Institutional research plan: CEZ:AV0Z50380511
Keywords : TRITICUM-AESTIVUM L. * HEXAPLOID WHEAT * BREAD WHEAT
Subject RIV: EB - Genetics ; Molecular Biology
Impact factor: 2.436, year: 2011

Positional cloning in bread wheat is a tedious task due to its huge genome size and hexaploid character. BAC libraries represent an essential tool for positional cloning. However, wheat BAC libraries comprise more than milion clones, which makes thein screening very laborious. Here, we present a targeted approach based on chromosome-specific BAC libraries. Such libraries were constructed from flow-sorted arms of wheat chromosome 7D. A library from the short arm (7DS) consisting of 49,152 clones with 113 kb insert size represented 12.1 arm equivalents whereas a library from the long arm (7DL) comprised 50,304 clones of 116 kb providing 14.9x arm coverage. The 7DS library was PCR screened with markers linked to Russian wheat aphid resistance gene DnCI2401, the 7DL library was screened by hybridization with a probe linked to greenbug resistence gene Gb3. The small number of clones combined with high coverage made the screening highly efficient and cost effective.
Permanent Link: http://hdl.handle.net/11104/0006543