Yeast microarrays for genome wide parallel genetic and gene expression analysis

Abstract

We have developed high-density DNA microarrays of yeast ORFs. These microarrays can monitor hybridization to ORFs for applications such as quantitative differential gene expression analysis and screening for sequence polymorphisms. Automated scripts retrieved sequence information from public databases to locate predicted ORFs and select appropriate primers for amplification. The primers were used to amplify yeast ORFs in 96-well plates, and the resulting products were arrayed using an automated micro arraying device. Arrays containing up to 2,479 yeast ORFs were printed on a single slide. The hybridization of fluorescently labeled samples to the array were detected and quantitated with a laser confocal scanning microscope. Applications of the microarrays are shown for genetic and gene expression analysis at the whole genome level.

Figure 1

Two-color fluorescent scan of a yeast microarray containing 2,479 elements (ORFs). The center-to-center distance between elements is 345 μm. A probe mixture consisting of cDNA from yeast extract/peptone (YEP) galactose (green pseudocolor) and YEP glucose (red pseudocolor) grown yeast cultures was hybridized to the array. Intensity per element corresponds to ORF expression, and pseudocolor per element corresponds to relative ORF expression between the two cultures.

Figure 2

ORF categories displaying differential expression between heat shocked and untreated cultures. Bars within categories correspond to individual ORFs. Green shaded bars correspond to relative increases in ORF expression under 25°C growth conditions. Red shaded bars correspond to relative increases in ORF expression under 39°C growth conditions.