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

Abstract

Abstract A stable internal control gene or house keepinggene (HKG) is often used to normalise mRNA levels indifferent samples for expression analysis. In the presentstudy, the authors identified and evaluated three HKGs,beta actin (b-actin), glyceraldehyde-3-phosphate dehydro-genase (GAPDH) and elongation factor 1 alpha (EF1a) forgene expression study in Catla (Catla catla). Geneexpression levels were quantified by quantitative real-timereverse transcription polymerase chain reaction in differenttissues (liver, kidney, intestine, gill, muscle and brain) anddevelopmental stages (0, 3, 6, 12, 24 h, and 5, 7, 9 dayspost-fertilization). Expression stability was evaluated bycomparing the coefficients of variation of the cyclethreshold values and stability index. All the tested HKGsexhibited wide expression range. The results showed thatb-actin is the most stable gene followed by the EF1aandGAPDH in different tissues whereas GAPDH was moststable gene followed by EF1aand b-actin during embry-onic development.