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First published online November 7, 2002; 10.1104/pp.012567

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Plant Physiol, December 2002, Vol. 130, pp. 1614-1625

Targeted Analysis of Orthologous Phytochrome A Regions of the Sorghum, Maize, and Rice Genomes using Comparative Gene-Island Sequencing1

Daryl T. Morishige, Kevin L. Childs, L. David Moore, and John E. Mullet*

Institute for Plant Genomics and Biotechnology and Department of Biochemistry and Biophysics, Texas A&M University, College Station, Texas 77843

A "gene-island" sequencing strategy has been developed that expedites the targeted acquisition of orthologous gene sequences from related species for comparative genome analysis. A 152-kb bacterial artificial chromosome (BAC) clone from sorghum (Sorghum bicolor) encoding phytochrome A (PHYA) was fully sequenced, revealing 16 open reading frames with a gene density similar to many regions of the rice (Oryza sativa) genome. The sequences of genes in the orthologous region of the maize (Zea mays) and rice genomes were obtained using the gene-island sequencing method. BAC clones containing the orthologous maize and rice PHYA genes were identified, sheared, subcloned, and probed with the sorghum PHYA-containing BAC DNA. Sequence analysis revealed that approximately 75% of the cross-hybridizing subclones contained sequences orthologous to those within the sorghum PHYA BAC and less than 25% contained repetitive and/or BAC vector DNA sequences. The complete sequence of four genes, including up to 1 kb of their promoter regions, was identified in the maize PHYA BAC. Nine orthologous gene sequences were identified in the rice PHYA BAC. Sequence comparison of the orthologous sorghum and maize genes aided in the identification of exons and conserved regulatory sequences flanking each open reading frame. Within genomic regions where micro-colinearity of genes is absolutely conserved, gene-island sequencing is a particularly useful tool for comparative analysis of genomes between related species.


1 This material is based on work supported by the National Science Foundation (grant no. 0077713), by the Texas Agricultural Experiment Station, and by the Perry Adkisson Chair in Agricultural Biology.

* Corresponding author; e-mail jmullet{at}tamu.edu; fax 979-862-4718.

© 2002 American Society of Plant Biologists



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