ABSTRACT
Functional analysis of a genome requires accurate gene structure information and a complete gene inventory. A dual experimental strategy was used to verify and correct the initial genome sequence annotation of the reference plant Arabidopsis. Sequencing full-length cDNAs and hybridizations using RNA populations from various tissues to a set of high-density oligonucleotide arrays spanning the entire genome allowed the accurate annotation of thousands of gene structures. We identified 5817 novel transcription units, including a substantial amount of antisense gene transcription, and 40 genes within the genetically defined centromeres. This approach resulted in completion of approximately 30% of the Arabidopsis ORFeome as a resource for global functional experimentation of the plant proteome.
Subject(s)
Arabidopsis/genetics , Genome, Plant , RNA, Messenger/genetics , RNA, Plant/genetics , Transcription, Genetic , Chromosome Mapping , Chromosomes, Plant/genetics , Cloning, Molecular , Computational Biology , DNA, Complementary/genetics , DNA, Intergenic , Expressed Sequence Tags , Gene Expression Profiling , Genes, Plant , Genomics , Nucleic Acid Hybridization , Oligonucleotide Array Sequence Analysis , Open Reading Frames , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
Over 225,000 independent Agrobacterium transferred DNA (T-DNA) insertion events in the genome of the reference plant Arabidopsis thaliana have been created that represent near saturation of the gene space. The precise locations were determined for more than 88,000 T-DNA insertions, which resulted in the identification of mutations in more than 21,700 of the approximately 29,454 predicted Arabidopsis genes. Genome-wide analysis of the distribution of integration events revealed the existence of a large integration site bias at both the chromosome and gene levels. Insertion mutations were identified in genes that are regulated in response to the plant hormone ethylene.
