Quantitative analysis of gender-regulated transcripts in the filarial nematode Brugia malayi by real-time RT-PCR

Molecular and Biochemical Parasitology
Ben-Wen LiGary Weil

Abstract

Improved understanding of the biology of reproduction in filarial worms may lead to identification of new targets for drugs or vaccines. Real-time RT-PCR is increasingly being adopted for RNA quantification and genetic analysis. Candidate gender-regulated genes were selected from genes identified in prior studies by differential display RT-PCR and by electronic selection of the Brugia malayi expression sequence tag (EST) database for clusters with possible gender-specific expression (four or more transcripts in male cDNA library ESTs but none in female ESTs or vice versa). Expression of candidate genes in male and female worms was compared by real-time reverse transcription PCR with sequence-specific primers. Double stranded DNA product was measured by SYBR Green I fluorescence; melting curves and agarose gel electrophoresis were used to verify the specificity of results. Relative gene expression results were normalized by parallel studies with internal control genes that were shown to be equally expressed in male and female worms (beta actin 2B, histone H3, NADH dehydroxygenase subunit 1) and calculated by the comparative C(t) method. Nineteen of 31 candidate genes were verified to have reproducible, gender-biased expression w...Continue Reading

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Citations

Mar 10, 2012·Parasitology Research·Sivasakthivel ThirugnanamKalyanasundaram Ramaswamy
Apr 11, 2013·Molecular Genetics and Genomics : MGG·Zhengyuan WangMakedonka Mitreva
Jun 12, 2009·Archives of Virology·Chan-Juan ShenDe-Kang Zhu
Nov 9, 2007·PLoS Neglected Tropical Diseases·Catherine BourguinatRoger K Prichard
Jul 24, 2012·International Journal for Parasitology·Daojun JiangBenwen Li
Jul 23, 2015·International Journal for Parasitology. Drugs and Drug Resistance·Ben-Wen LiGary J Weil
Jan 25, 2008·Molecular and Biochemical Parasitology·Catherine BourguinatRoger K Prichard
Jan 10, 2008·Trends in Parasitology·Alasdair J NisbetRobin B Gasser
Nov 16, 2007·International Journal for Parasitology·Daojun JiangGary J Weil
Jul 11, 2006·Molecular and Biochemical Parasitology·Ben-Wen LiMakedonka Mitreva
Sep 20, 2005·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Paul T Monis, Steven Giglio
Jul 5, 2005·Molecular and Biochemical Parasitology·Shweta Trivedi, Prema Arasu
Apr 17, 2016·International Journal of Cardiology·Zuoguang WangYongxiang Wei
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Aug 16, 2005·Trends in Genetics : TIG·Makedonka MitrevaJames P McCarter

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