A sunflower BAC library suitable for PCR screening and physical mapping of targeted genomic regions.
A sunflower BAC library consisting of 147,456 clones with an average size of 118 kb has been constructed and characterized. It represents approximately 5x sunflower haploid genome equivalents. The BAC library has been arranged in pools and superpools of DNA allowing screening with various PCR-based markers. Each of the 32 superpools contains 4,608 clones and corresponds to a 36 matrix pools. Thus, the screening of the entire library could be accomplished in less than 80 PCR reactions including positive and negative controls. As a demonstration of the feasibility of the concept, a set of 24 SSR markers covering about 36 cM in the sunflower SSR map (Tang et al. in Theor Appl Genet 105:1124-1136, 2002) have been used to screen the BAC library. About 125 BAC clones have been identified and then organized in 23 contigs by HindIII digestion. The contigs are anchored on the SSR map and thus constitutes a first-generation physical map of this region. The utility of this BAC library as a genomic resource for physical mapping and map-based cloning in sunflower is discussed.
Bouzidi
Mohamed Fouad
MF
UMR 1095 INRA-UBP Amélioration et Santé des Plantes, Université Blaise Pascal, 24 avenue des Landais, 63177, Aubière Cedex, France.
Franchel
Jérôme
J
Tao
Quanzhou
Q
Stormo
Keith
K
Mraz
A
A
Nicolas
Paul
P
Mouzeyar
Saïd
S
D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D003001:Cloning, Molecular; D016172:DNA Fingerprinting; D018744:DNA, Plant / Q000235:genetics; D015723:Gene Library; D005819:Genetic Markers; D018745:Genome, Plant; D006368:Helianthus / Q000235:genetics*; D018598:Minisatellite Repeats; D020161:Physical Chromosome Mapping; D016133:Polymerase Chain Reaction
D018744:DNA, Plant; D005819:Genetic Markers