Evaluation of housekeeping genes as references for quantitative real-time PCR analysis of gene expression in the murrel Channa striatus under high-temperature stress.
Quantitative real-time polymerase chain reaction is the most advanced method of quantifying gene expression studies; however, the significance of the obtained results strongly depends on the normalization of the data to compensate for differences between the samples. In the present study, expression analysis of six different constitutively expressed genes viz. 18S ribosomal RNA, glyceraldehyde-3-phosphate dehydrogenase (gapdh), beta actin (βactin), ribosomal binding protein L13, tubulin and TATA-box-binding protein (tbp) were carried out to test their efficacy as reference genes in three different tissues, namely liver, gill and muscle of murrel Channa striatus exposed to high temperature for variable time periods. The stability and suitability of the genes were determined by using bioinformatic tools: GeNorm, NormFinder and BestKeeper. Based on the results, tub/βactin could be used as the reference genes for liver and gill tissues and βactin/gapdh could be the reference genes for muscle tissues in Channa striatus under both short- and long-term thermal stress.
KIIT School of Biotechnology, KIIT University, Bhubaneswar, Odisha, 751024, India.
Gopal Krishna
GK
Purohit
Fishery Resource and Environmental Management Division, ICAR- Central Inland Fisheries Research Institute, Barrackpore, Kolkata, 700120, India.
Arabinda
A
Mahanty
Fishery Resource and Environmental Management Division, ICAR- Central Inland Fisheries Research Institute, Barrackpore, Kolkata, 700120, India. bimalmohanty12@rediffmail.com.
Bimal Prasanna
BP
Mohanty
KIIT School of Biotechnology, KIIT University, Bhubaneswar, Odisha, 751024, India. smohanty@kiitbiotech.ac.in.
Sasmita
S
Mohanty
D000199:Actins / Q000235:genetics; D000818:Animals; D029941:Fish Proteins / Q000235:genetics*; D015870:Gene Expression; D020043:Genes, Essential / Q000235:genetics*; D005880:Gills / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D006358:Hot Temperature / Q000009:adverse effects*; D008099:Liver / Q000378:metabolism; D009132:Muscles / Q000378:metabolism; D010473:Perciformes / Q000235:genetics* / Q000502:physiology; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D012269:Ribosomal Proteins / Q000235:genetics; D013312:Stress, Physiological / Q000235:genetics*; D035181:TATA-Box Binding Protein / Q000235:genetics; D014404:Tubulin / Q000235:genetics
D000199:Actins; D029941:Fish Proteins; D012337:RNA, Ribosomal, 18S; D012269:Ribosomal Proteins; D035181:TATA-Box Binding Protein; D014404:Tubulin; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases
Channa striatus; Normalization; Quantitative real-time PCR; Reference genes; Thermal stress