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 million clones, which makes their 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 resistance gene Gb3. The small number of clones combined with high coverage made the screening highly efficient and cost effective.
- MeSH
- chromozomy rostlin genetika MeSH
- fluorescence MeSH
- hybridizace nukleových kyselin genetika MeSH
- karyotypizace MeSH
- klonování DNA metody MeSH
- mikrosatelitní repetice genetika MeSH
- mšice fyziologie MeSH
- nemoci rostlin genetika imunologie parazitologie MeSH
- polymerázová řetězová reakce MeSH
- přirozená imunita genetika MeSH
- pšenice genetika imunologie parazitologie MeSH
- rostlinné geny genetika MeSH
- umělé bakteriální chromozomy genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
