mRNA EXPRESSIONPROFILINGOF GLUTATHIONE PEROXIDASE (GPX GENE) USING FIVE MEDICINAL PLANTS ON S. CEREVISIAEAbstract
Glutathione peroxidase gene expression profiling has been determined using budding yeast, S. cerevisiae, as a model organism mainly used for extensively reliable microbial studies of various cellular processes evolved in evolutionarily distant species. Roots of five medicinal plants were used: Withania somnifera, Terminalia arjuna, Ranunculus sceleratus, Bacopa monnieri and Acalypha indica. The maceration method exhibited the extraction process, followed by S. cerevisiae culturing at 600nm. Classic Trizol method for RNA extraction of yeast cells. Further, evaluation of the quantity and quality of the isolated RNA before the CDNA synthesis, respectively. Polymerase Chain Reaction was performed using specific conditions, having 60OC as GPx primers annealing temperature. Real-Time Polymerase Chain Reaction of samples was performed where Beta-actin primers as a housekeeping gene. High purity of RNA samples was yielded to be in the range of 1.8-2.0. The size of cDNA PCR products was found to be 118bp. The RT-qPCR results showed high over-expression, i.e., fold change unit of GPx gene in all sample extracts compared to the control yeast. Ethanol and methanol extracts of Acalypha indica, ethanol extract of Ranunculus sceleratus, and methanol extract of Terminaliya arjuna showed high overexpression of GPx gene. The fold change of GPxgene profiling ranged from 17.638 ± 0.1-64.415 ± 0.18. One-way ANOVA was calculated, showing p<0.05 considerably as highly significant. Therefore, the bioactive components of five plant root extracts can potentially enhance the antioxidant GPX gene expression in S. cerevisiae at an extensively good level.
Debanjana Prasad and Shailesh Solanki *
Department of Agriculture and Environment, Noida International University, Greater Noida, Uttar Pradesh, India.
01 July 2022
11 August 2022
31 August 2022
01 March 2023