The Effects of Zinc on Gene Expression in Saccharomyces cerevisiae using Polymerase Chain Reaction (PCR) for Six Candidate Genes

The Effects of Zinc on Gene Expression in Saccharomyces cerevisiae using Polymerase Chain Reaction (PCR) for Six Candidate Genes

In this experiment the effects of various concentrations of zinc on Saccharomyces cerevisiae will be investigated using Polymerase Chain Reaction (PCR). Zinc (Zn) is one of the most naturally occurring elements. It is an important integral cofactor for many cellular activities. Zinc participates in all major biochemical reactions such as transcription of DNA and translation of RNA. More than three hundred enzymes require zinc for their catalytic function. Saccharomyces cerevisiae strain S288C is a species of budding yeast. It is the most intensively studied eukaryotic model in molecular and cell biology. The mRNA from these yeast cells grown in different concentrations of zinc (0μM, 1μM, 50μM, 1mM) will be reviewed to determine the expression of patterns specific genes. In particular, the expression of six genes ZRT1, ZRT2, ZRT3, ZRC1, ZAP1, and VEL1 will be examined. The yeast will be grown in CSD media with the appropriate concentration of zinc. Zinc will be added as zinc chloride. A control sample will be grown without the addition of zinc chloride. The 1mM zinc is considered to be an excess amount, while 1μM is considered to be less than the minimum required Zn concentration. RNA will be isolated from yeast exposed to each environmental condition. The mRNA from the total RNA extracted will then be converted to cDNA, and the presence of the various genes will be tested using specially selected primers and polymerase chain reaction (PCR).