Quantitative real-time PCR analysis of Anopheles dirus TEP1 and NOS during Plasmodium berghei infection, using three reference genes

Quantitative reverse transcription PCR (qRT-PCR) has been an integral part of characterizing the immunity of mosquitoes towards invasion. Two anti- factors of , thioester-containing protein 1 (TEP1) and nitric oxide synthase (NOS), play a role in the refractoriness of towards infection and are generally expressed during infection. However, these are less studied in , a dominant malaria vector in Southeast Asia. Furthermore, most studies used a single reference gene for normalization during gene expression analysis without proper validation. This may lead to erroneous quantification of expression levels. Therefore, the present study characterized and investigated the expression profiles of and of during infection Prior to that, the ( ), ( ) and ( ) genes were validated for their suitability as a set of reference genes. and expressions in were found to be significantly induced after infection.