Effect OF M. koenigii on the expression of cell wall formation related genes (mecA and fmhb)

Abstract

Introduction

Generally, quantitative polymerase-chain-reaction technique (qPCR) has been recognized as the gold standard for gene expression analysis. It supersedes the original conventional PCR, due to its ability of measuring the amplification of cDNA in real time as the reaction progresses.

Aims

The aim of this study was to determine the effects of the ethyl acetate leaf extract of Murraya koenigii (M. koenigii) on the expression of cell wall formation related genes (mecA and fmhb).

Materials and Methods

Ribonucleic acid (RNA) of the bacterial cells (Staphylococcus aureus ATCC 700,699) was extracted using Trizol reagent. The concentrations and purities of all RNA analysis were obtained from NanoDrop Spectrophotometer. cDNA Synthesis Kit was used for cDNA synthesis. The integrity of the cDNA was identified using ethidium bromide, through 1.5% agarose gel electrophoresis in 1 x TBE buffer. Finally, quantitative real-time PCR technique was employed to establish and validate the antibacterial activity of the plant extract on gene expression of the selected genes at the cellular level and the quantification of the gene's expression was determined using delta-delta Ct method.

Result

The result revealed that the exposure of the bacterial cells to the plant extract instigated upregulation of the selected genes. This indicates resistance of the bacteria to the treated extract against the selected cell wall formation genes

Conclusion

These findings suggest that the ethyl acetate leaf extract of M. koenigii lacks potential antibacterial activity on the expression of cell wall formation related genes (mecA and fmhb) of S. aureus bacteria.