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

PeerJ

article

Jonathan W.K. Liew¹  Mun Yik Fong¹  Yee Ling Lau¹ 

[1] Department of Parasitology, Faculty of Medicine, University of Malaya

关键词: Anopheles dirus;    Plasmodium berghei;    TEP1;    NOS;    Reference genes;    Normalization;   

DOI  :  10.7717/peerj.3577

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

Quantitative reverse transcription PCR (qRT-PCR) has been an integral part of characterizing the immunity of Anopheles mosquitoes towards Plasmodium invasion. Two anti-Plasmodium factors of Anopheles, thioester-containing protein 1 (TEP1) and nitric oxide synthase (NOS), play a role in the refractoriness of Anopheles towards Plasmodium infection and are generally expressed during infection. However, these are less studied in Anopheles dirus, 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 TEP1 and NOS of Anopheles dirus during P. berghei infection. Prior to that, the elongation factor 1-alpha (EF1), actin 1 (Act) and ribosomal protein S7 (S7) genes were validated for their suitability as a set of reference genes. TEP1 and NOS expressions in An. dirus were found to be significantly induced after P. berghei infection.

【 授权许可】

CC BY