INVESTIGADORES
YANOVSKY Marcelo Javier
artículos
Título:
Rapid Array Mapping of Circadian Clock and Developmental Mutations in Arabidopsis.
Autor/es:
? SP HAZEN, JO BOREVITZ, FG HARMON, JL PRUNEDA-PAZ, TF SCHULTZ, MJ YANOVSKY, SJ LILJEGREN, JR ECKER, AND SA KAY
Revista:
PLANT PHYSIOLOGY.
Referencias:
Año: 2005 p. 990 - 997
ISSN:
0032-0889
Resumen:
Classical forward genetics, the identification of genes responsible for mutant phenotypes, remains an important part of
functional characterization of the genome. With the advent of extensive genome sequence, phenotyping and genotyping
remain the critical limiting variables in the process of map-based cloning. Here, we reduce the genotyping problem by
hybridizing labeled genomic DNA to the Affymetrix Arabidopsis (Arabidopsis thaliana) ATH1 GeneChip. Genotyping was
carried out on the scale of detecting greater than 8,000 single feature polymorphisms from over 200,000 loci in a single assay. By
combining this technique with bulk segregant analysis, several high heritability development and circadian clock traits were
mapped. The mapping accuracy using bulk pools of 26 to 100 F2 individuals ranged from 0.22 to 1.96 Mb of the mutations
revealing mutant alleles of EARLY FLOWERING 3, EARLY FLOWERING 4, TIMING OF CAB EXPRESSION 1, andArabidopsis thaliana) ATH1 GeneChip. Genotyping was
carried out on the scale of detecting greater than 8,000 single feature polymorphisms from over 200,000 loci in a single assay. By
combining this technique with bulk segregant analysis, several high heritability development and circadian clock traits were
mapped. The mapping accuracy using bulk pools of 26 to 100 F2 individuals ranged from 0.22 to 1.96 Mb of the mutations
revealing mutant alleles of EARLY FLOWERING 3, EARLY FLOWERING 4, TIMING OF CAB EXPRESSION 1, and2 individuals ranged from 0.22 to 1.96 Mb of the mutations
revealing mutant alleles of EARLY FLOWERING 3, EARLY FLOWERING 4, TIMING OF CAB EXPRESSION 1, andEARLY FLOWERING 3, EARLY FLOWERING 4, TIMING OF CAB EXPRESSION 1, and
ASYMMETRIC LEAVES 1. While direct detection of small mutations, such as an ethyl-methane sulfonate derived single
base substitutions, is limited by array coverage and sensitivity, large deletions such as those that can be caused by fast neutrons
are easily detected. We demonstrate this by resolving two deletions, the 77-kb flavin-binding, kelch repeat, f-box 1 and the 7-kb
cryptochrome2-1 deletions, via direct hybridization of mutant DNA to ATH1 expression arrays.. While direct detection of small mutations, such as an ethyl-methane sulfonate derived single
base substitutions, is limited by array coverage and sensitivity, large deletions such as those that can be caused by fast neutrons
are easily detected. We demonstrate this by resolving two deletions, the 77-kb flavin-binding, kelch repeat, f-box 1 and the 7-kb
cryptochrome2-1 deletions, via direct hybridization of mutant DNA to ATH1 expression arrays.flavin-binding, kelch repeat, f-box 1 and the 7-kb
cryptochrome2-1 deletions, via direct hybridization of mutant DNA to ATH1 expression arrays.ryptochrome2-1 deletions, via direct hybridization of mutant DNA to ATH1 expression arrays.