Abstract
Housekeeping gene is a gene expressed with a fixed level and in abundant amounts under various conditions. After validation, the housekeeping gene can be used as an internal control to normalize gene expression data. This study reports the isolation of several housekeeping genes in Durik-durik plant (Syzygium sp). This plant material in form of fresh leaves from Durik-durik plants are taken from Kajuik Lake, Riau Province. The next stage is total DNA isolation, polymerase chain reaction, electrophoresis, sequencing and data analysis using bioinformatic tools. The isolated housekeeping genes included 18S rRNA, actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Beta-tubulin and Ubiquitin with sizes of 400 bp, 679 bp, 1134 bp, 836 bp, 1167 bp and 2155 bp, respectively. In addition to 18S rRNA, the five housekeeping genes are the first reported from the genus Syzygium and referable to isolate housekeeping genes in other species in this genus. The six housekeeping genes can be used as internal controls on Durik-durik plants after validation.
Highlights
Gene expression studies need some internal controls that commonly come from housekeeping genes group
The Polymerase Chain Reaction (PCR) products obtained in encoding genes of 18S rRNA (Syz-18S), actin (Syz-ACT), glyceraldehyde-3-phosphate dehydrogenase (SyzGAPDH), elongation factor 1-alpha (Syz-EF1α), beta-tubulin Syz-βTUB) and ubiquitin
The identity and query cover valueswere very high on the BLASTn analysis based on genes encoding the 18S rRNA, actin, EF1α and β-tubulin, ranging from 86%-99% and 80%100%, respectively
Summary
Gene expression studies need some internal controls that commonly come from housekeeping genes group. Housekeeping genes are group of genes expressed constitutively at a constant level under various conditions This gene group encodes the proteins involved in the various metabolisms that are essential for the survival of a cell (Koonin, 2000). Actin and tubulin proteins are involved in cytoskeleton formation because they play a role in maintaining cellular turgidity and structure (Keren et al, 2008; Kueh & Mitchison, 2009). Another example is the ubiquitin protein involved in the recycling of cell components through the ubiquitination process (Callis, 2014; Stone, 2014). Analysis of evolution of exon– intron structure of eukaryotic genes.
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