Detection of single copy sequences using BAC-FISH and C-PRINS techniques in sunflower chromosomes
Biocell
; 35(1): 19-28, Apr. 2011. ilus, tab
Article
en En
| LILACS
| ID: lil-595006
Biblioteca responsable:
AR40.1
ABSTRACT
Bacterial artificial chromosome-fluorescence in situ hybridization (BAC-FISH) and cycling-primed in situ labeling (C-PRINS) techniques were evaluated for integration of physical and genetic maps of sunflower (Helianthus annuus L.). Single-site SSR markers were selected from three linkage groups of a high-density sunflower genetic map. This selection was based on previously identified QTL associated to S. sclerotiorum. These markers were used to select BACs contaning single copy sequences for BAC-FISH aplication. Blocking of highly dispersed repetitive sunflower sequences reduced unspecific hybridization, and allowed the detection of specific signals for BACs containing SSR markers HA4222 and HA2600, anchored to LG 16 and LG 10, respectively. Single-site FISH signal detection was optimized by adjusting the relative quantity and quality of unlabelled repetitive sequences present in the blocking DNA. The SSR marker ORS1247 anchored to the LG 17 was detected by C-PRINS, which yielded fluorescence signals that were specific and intense. This progress in localizing single-copy sequences using BAC-FISH and indirect C-PRINS strategies in sunflower will facilitate the integration of genetic and physical maps, allowing the identification of chromosomes containing key genes and/or QTL associated to agronomic important traits in sunflower.
Texto completo:
1
Índice:
LILACS
Asunto principal:
Análisis de Secuencia de ADN
/
Hibridación Fluorescente in Situ
/
Cromosomas Artificiales Bacterianos
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Cromosomas de las Plantas
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Helianthus
Tipo de estudio:
Diagnostic_studies
/
Prognostic_studies
Idioma:
En
Revista:
Biocell
Asunto de la revista:
Clulas
Año:
2011
Tipo del documento:
Article