Characterisation and validation of housekeeping gene for expression analysis in Catla catla (Hamilton).

Abstract

A stable internal control gene or house keeping gene (HKG) is often used to normalise mRNA levels in different samples for expression analysis. In the present study, the authors identified and evaluated three HKGs, beta actin (β-actin), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and elongation factor 1 alpha (EF1α) for gene expression study in Catla (Catla catla). Gene expression levels were quantified by quantitative real-time reverse transcription polymerase chain reaction in different tissues (liver, kidney, intestine, gill, muscle and brain) and developmental stages (0, 3, 6, 12, 24 h, and 5, 7, 9 days post-fertilization). Expression stability was evaluated by comparing the coefficients of variation of the cycle threshold values and stability index. All the tested HKGs exhibited wide expression range. The results showed that β-actin is the most stable gene followed by the EF1α and GAPDH in different tissues whereas GAPDH was most stable gene followed by EF1α and β-actin during embryonic development.