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

Zinc (Zn) is one of the principle trace elements required in biological systems, with structural or enzymatical roles in hundreds of proteins. In humans, zinc deficiency has gradually come to be recognized as a clinically significant and common form of malnutrition, particularly in chronically ill patients. The clinical manifestations of zinc deficiency are diverse with effects on the immune system, appetite, and embryonic development. The regulation of zinc distribution remains a critical, unanswered question. The manner in which zinc is released from its tight binding sites in proteins and its transfer from one site to another are also unknown. In this study, microarray technology was used to examine the potential effects of Zn exposure (0μM, 1μM, 50μM, 1mM Zn solutions for 24 hours) on gene expression in Saccharomyces cerevisiae (yeast), a model organism that shares roughly 31% of its genome with humans. However, due to technical complications, the microarray images were inconclusive due to inadequate signal binding. As an alternative approach, the expression of four genes was examined using reverse transcriptase-polymerase chain reaction (RT-PCR). The genes selected were IZH1, IZH4, VEL1 and TDH1. IZH4 was expected to be induced in high zinc conditions. IZH1 was expected to be repressed in low zinc conditions. TDH1 is a gene directly involved in metabolism and was expected to be expressed in all zinc conditions. VEL1, on the other hand, is a gene of unknown function, thought to be involved in velum formation and was expected to be expressed in low zinc conditions.