THE CHLAMYDOMONAS GENOME PROJECT

Grossman, A. R. 1 , Davies, J. 2 , Shrager, J. 1 , Chang, C.‐W 1 , McDermott, J. 3 & Zhang, Z. 11 Department of Plant Biology, The Carnegie Institution of Washington, 260 Panama Street, Stanford, CA 94305 USA; 2 Exelixis Pharmaceuticals, Inc. 170 Harbor Way, So. San Francisco, CA 94083‐051 USA; 3 Department of Botany, Iowa State University, Ames, IA 50011‐1020 USA The Chlamydomonas genome project involves 1) sequencing of cDNAs isolated from cells exposed to various environmental conditions, 2) construction of a high density DNA microarray, 3) construction of genomic contigs that nucleate around specific physical and genetic markers, 4) generation of a complete chloroplast genome sequence and analyses of chloroplast gene expression, and 5) placement of the genomic information on the network in a user friendly format. The aspects of the project emphasized by our group at Stanford involves the generation of normalized cDNA libraries, sequencing of the cDNAs to generate both contigs and unigene families and the use of this information to construct a high density cDNA microarray. I will discuss the techniques involved in securing cDNA sequence information and the ways in which that information is assembled and analyzed. I will also discuss the use of the different cDNA libraries to identifying differentially expressed genes (by in silico subtractions) and strategies for constructing high‐density cDNA microarrays. Finally, if completed, the first global expression studies, and the use of the microarrays to elucidate global expression in mutant strains will be discussed.