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

PeerJ. 2017;5:e3577 DOI 10.7717/peerj.3577

 

Journal Homepage

Journal Title: PeerJ

ISSN: 2167-8359 (Online)

Publisher: PeerJ Inc.

LCC Subject Category: Medicine

Country of publisher: United States

Language of fulltext: English

Full-text formats available: PDF, HTML, XML

 

AUTHORS

Jonathan W.K. Liew (Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia)
Mun Yik Fong (Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia)
Yee Ling Lau (Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia)

EDITORIAL INFORMATION

Blind peer review

Editorial Board

Instructions for authors

Time From Submission to Publication: 10 weeks

 

Abstract | Full Text | Full Text

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.