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Título : Validation of housekeeping genes as an internal control for gene expression studies in Giardia lamblia using quantitative real-time PCR
Creador: Marcial Quino Jaime
Nivel de acceso: Open access
Palabras clave : Animales
Antiprotozoarios - farmacología
Cartilla de ADN
Regulación de la Expresión Génica
Genes Esenciales
Giardia lamblia - Efectos de drogas
Giardia lamblia - genética
Metronidazol - farmacología
Reacción en Cadena en Tiempo Real de la Polimerasa
Tiazoles - farmacología
Animals
Antiprotozoal Agents - pharmacology
DNA Primers
Gene Expression Regulation
Genes, Essential
Giardia lamblia - drug effects
Giardia lamblia - genetics
Metronidazole - pharmacology
Real-Time Polymerase Chain Reaction
Thiazoles - pharmacology
G. lamblia; Expresión del gen; Genes de limpieza; RT-qPCR; Programa RefFinder; Validación
G. lamblia; Gene expression; Housekeeping genes; RT-qPCR; RefFinder program; Validation
Descripción : The analysis of transcript levels of specific genes is important for understanding transcriptional regulation and for the characterization of gene function. Real-time quantitative reverse transcriptase PCR (RT-qPCR) has become a powerful tool to quantify gene expression. The objective of this study was to identify reliable housekeeping genes in Giardia lamblia. Twelve genes were selected for this purpose, and their expression was analyzed in the wild type WB strain and in two strains with resistance to nitazoxanide (NTZ) and metronidazole (MTZ), respectively. RefFinder software analysis showed that the expression of the genes is different in the three strains. The integrated data from the four analyses showed that the NADH oxidase (NADH) and aldolase (ALD) genes were the most steadily expressed genes, whereas the glyceraldehyde-3-phosphate dehydrogenase gene was the most unstable. Additionally, the relative expression of seven genes were quantified in the NTZ- and MTZ-resistant strains by RT-qPCR, using the aldolase gene as the internal control, and the results showed a consistent differential pattern of expression in both strains. The housekeeping genes found in this work will facilitate the analysis of mRNA expression levels of other genes of interest in G. lamblia. © 2016 Elsevier B.V..
Colaborador(es) u otros Autores: Fierro Francisco
De La Mora-De La Mora Ignacio
Enríquez-Flores Sergio
Gómez-Manzo Saúl
Vanoye-Carlo America
Garcia-Torres Itzhel
Sierra-Palacios Edgar
Reyes-Vivas Horacio
Fecha de publicación : 2016
Tipo de publicación: Artículo
Formato: pdf
Identificador del Recurso : 10.1016/j.gene.2016.01.018
Fuente: Gene 581(1):21 - 30
URI : http://repositorio.pediatria.gob.mx:8180/handle/20.500.12103/2948
Idioma: eng
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