Microarray Analysis of the ZMS2 Transcription Factor

Kristen Caldwell, Peggy Kipa, and Nhut Le

Abstract:

Oxidative stress can harm cells by damaging DNA and proteins.  G6PD is a protein activated by yeast cells in response to oxidative stress and is encoded by the ZWF1 gene.  Upregulation of the genes ZMS1 and ZMS2 has been shown to suppress the ZWF1 deletion under conditions of oxidative stress.  DNA microarrays were used to study genes expression in ZMS2⁺⁺ yeast strains.  Isolated RNA was reversed transcribed into cDNA, hybridized to DNA chips, and tagged with fluorescent dyes.  The resulting microarray yielded no usable data.  Microarrays from Fall 2004 were used for data analysis.  From these arrays, four genes that used the same transcription factors were found to be consistently upregulated between arrays.  Two genes were found to be consistently under expressed between arrays.  Future research could explore the function of these genes.  Also, other microarray protocols could be used to definitively determine the genes affected by the potential ZMS2 transcription factor.

Individual Projects:

Four common over expressed genes among four different microarray slides from the Fall 2004 data were found.  Their promoter sequences and function were found using the Yeast Genome Database.  Similar transcription factors of the four genes were found using the YEASTRACT website.

The most over expressed and under expressed genes were determined of the ZMS1ΔZMS2Δ slide from the Fall 2004 data.  The genes were compared with another slide to see if any were common among both slides.  The functions of those genes were also compared.

Other methods for dyeing the cDNA were examined.  An alternative protocol is proposed for the microarray project using single channel arrays, as well as a proposal for obtaining cDNA immediately after RNA extraction.

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