Selection of reference genes for mRNA quantification in Trypanosoma brucei

被引：75

作者：

Brenndoerfer, Martin ^{[1
]}

Boshart, Michael ^{[1
]}

机构：

[1] Univ Munich, Dept Biol 1, Biozentrum, D-82152 Martinsried, Germany

来源：

MOLECULAR AND BIOCHEMICAL PARASITOLOGY | 2010年 / 172卷 / 01期

关键词：

mRNA expression; Normalization; Real time PCR; Reference gene; geNorm; Northern blot; TIME RT-PCR; BLOOD-STREAM FORMS; EXPRESSION ANALYSIS; DIFFERENTIATION; VALIDATION; NORMALIZATION; TRANSMISSION; SHOCK; LIFE; FLY;

D O I：

10.1016/j.molbiopara.2010.03.007

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Internal normalization is an established procedure that is necessary for accurate and reliable quantification of differentially regulated mRNAs. The profound changes of gene expression in parasitic life cycles pose a particular challenge on selection of appropriate reference genes for normalization, most importantly when using quantitative real time PCR (qPCR). Here we use the ranking algorithm implemented in the geNorm application to identify suitable Trypanosoma brucei reference genes for comparisons between the bloodstream and procyclic developmental stages and for analysis of mRNA induction by environmental conditions. For these conditions, the TERT gene is a good choice for valid normalization of qPCR and is clearly superior to some other reference genes reported in the literature. For comparison of other conditions, the ranking algorithm is recommended to verify a reliable and valid normalization that is instrumental to quantitative analysis of gene expression. (C) 2010 Elsevier B.V. All rights reserved.

引用

页码：52 / 55

页数：4

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