Identification of Housekeeping Genes of Mentha piperita

Abstract

Most of the housekeeping genes (HKGs) have constitutive expression in cells of living organisms. Their expression ensures the viability of the living organisms because HKGs control basic life processes of the cells. Expression of these genes is essential in all conditions even under biotic or abiotic stresses and is thought to be stable under any experimental conditions. Therefore, these genes can be used as reference genes or internal controls for the normalizations of the expression data under stressed conditions in comparisons to normal conditions. Recent studies showed that the expression of these reference genes can vary under different experimental conditions and no reference gene is perfect for all given treatments until properly validated. Most of the available information of such genes is from the economically important plants or model plants. This study reports the identification and partial sequencing of homologue Ubiquitin 10 gene of Mentha piperita. It has culinary uses. Leaves of this plant are used as raw salad and for making mint sauce and tea. Daily foods, confectionary, toffees and gums are mint flavored. It is now being used in manufacturing of cosmetics like shampoos, bath soaps, facials, creams, and perfumes. I designed gene specific primers of Ubiquitin 10 gene and amplified the genomic DNA extracted from leaves of Mentha piperita Amplified band of ~417 bp of Ubiquitin 10 gene was excided from the agarose gel and sequenced by using Beckman CEQ 8800 sequencer. BLAST was used to check the similarity index with other Ubiquitin 10 genes from other plants by using NCBI database to check its homology other Ubiquitin 10 gene from different plants. Similarly, Clustalw was used for DNA alignment and phylogenetic tree was constructed. It showed highly conserved nature of Ubiquitin 10 gene of M. piperita from different other plants. This gene can be used for the normalizations of the expression analysis by using reverse transcriptase Polymerase Reaction (RT-PCR) or quantitative real-time PCR data of this plant to explore several underlying mechanisms unique to its medicinal values.