Generation of an oligonucleotide array for analysis of gene expression in Chlamydomonas reinhardtii.
The availability of genome sequences makes it possible to develop microarrays that can be used for profiling gene expression over developmental time, as organisms respond to environmental challenges, and for comparison between wild-type and mutant strains under various conditions. The desired characteristics of microarrays (intense signals, hybridization specificity and extensive coverage of the transcriptome) were not fully met by the previous Chlamydomonas reinhardtii microarray: probes derived from cDNA sequences (approximately 300 bp) were prone to some nonspecific cross-hybridization and coverage of the transcriptome was only approximately 20%. The near completion of the C. reinhardtii nuclear genome sequence and the availability of extensive cDNA information have made it feasible to improve upon these aspects. After developing a protocol for selecting a high-quality unigene set representing all known expressed sequences, oligonucleotides were designed and a microarray with approximately 10,000 unique array elements (approximately 70 bp) covering 87% of the known transcriptome was developed. This microarray will enable researchers to generate a global view of gene expression in C. reinhardtii. Furthermore, the detailed description of the protocol for selecting a unigene set and the design of oligonucleotides may be of interest for laboratories interested in developing microarrays for organisms whose genome sequences are not yet completed (but are nearing completion).
Eberhard
Stephan
S
Department of Plant Biology, The Carnegie Institution, 260 Panama Street, Stanford, CA, 94305, USA. Stephan.Eberhard@ibpc.fr
Jain
Monica
M
Im
Chung Soon
CS
Pollock
Steve
S
Shrager
Jeff
J
Lin
Yuan
Y
Peek
Andrew S
AS
Grossman
Arthur R
AR
D000818:Animals; D002467:Cell Nucleus / Q000235:genetics; D016825:Chlamydomonas reinhardtii / Q000235:genetics* / Q000378:metabolism; D030561:Databases, Nucleic Acid; D020869:Gene Expression Profiling; D017125:Genes, Protozoan; D018503:Genome, Protozoan; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D013455:Sulfur / Q000172:deficiency / Q000378:metabolism
D013455:Sulfur