GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues

被引:638
作者
Barber, RD [1 ]
Harmer, DW [1 ]
Coleman, RA [1 ]
Clark, BJ [1 ]
机构
[1] Pharmagene Labs Ltd, Royston SG8 5HD, Herts, England
关键词
glyceraldehyde-3-phosphate dehydrogenase; gene expression; quantitative real-time reverse transcription-polymerase chain reaction;
D O I
10.1152/physiolgenomics.00025.2005
中图分类号
Q2 [细胞生物学];
学科分类号
071009 [细胞生物学]; 090102 [作物遗传育种];
摘要
Quantitative gene expression data are often normalized to the expression levels of control or so-called "housekeeping" genes. An inherent assumption in the use of housekeeping genes is that expression of the genes remains constant in the cells or tissues under investigation. Although exceptions to this assumption are well documented, housekeeping genes are of value in fully characterized systems. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is one of the most commonly used housekeeping genes used in comparisons of gene expression data. To investigate the value of GAPDH as a housekeeping gene in human tissues, the expression of GAPDH mRNA was measured in a panel of 72 different pathologically normal human tissue types. Measurements were obtained from 371,088 multiplexed, quantitative real-time RT-PCRs with specific target genes. Significant differences in the expression levels of GAPDH mRNA were observed between tissue types and between donors of the same tissue. A 15-fold difference in GAPDH mRNA copy numbers was observed between the highest and lowest expressing tissue types, skeletal muscle and breast, respectively. No specific effect of either age or gender was observed on GAPDH mRNA expression. These data provide an extensive analysis of GAPDH mRNA expression in human tissues and confirm previous reports of the marked variability of GAPDH expression between tissue types. These data establish comparative levels of expression and can be used to add value to gene expression data in which GAPDH is used as the internal control.
引用
收藏
页码:389 / 395
页数:7
相关论文
共 14 条
[1]
Validation of housekeeping genes for normalizing RNA expression in real-time PCR [J].
Dheda, K ;
Huggett, JF ;
Bustin, SA ;
Johnson, MA ;
Rook, G ;
Zumla, A .
BIOTECHNIQUES, 2004, 37 (01) :112-+
[2]
Validation of endogenous controls for gene expression studies in human adipocytes and preadipocytes [J].
Gorzelniak, K ;
Janke, J ;
Engeli, S ;
Sharma, AM .
HORMONE AND METABOLIC RESEARCH, 2001, 33 (10) :625-627
[3]
The use of quantitative RT-PCR to measure mRNA expression in a rat model of focal ischemia - caspase-3 as a case study [J].
Harrison, DC ;
Medhurst, AD ;
Bond, BC ;
Campbell, CA ;
Davis, RP ;
Philpott, KL .
MOLECULAR BRAIN RESEARCH, 2000, 75 (01) :143-149
[4]
A compendium of gene expression in normal human tissues [J].
Hsiao, LL ;
Dangond, F ;
Yoshida, T ;
Hong, R ;
Jensen, RV ;
Misra, J ;
Dillon, W ;
Lee, KF ;
Clark, KE ;
Haverty, P ;
Weng, ZP ;
Mutter, GL ;
Frosch, MP ;
MacDonald, ME ;
Milford, EL ;
Crum, CP ;
Bueno, R ;
Pratt, RE ;
Mahadevappa, M ;
Warrington, JA ;
Stephanopoulos, G ;
Stephanopoulos, G ;
Gullans, SR .
PHYSIOLOGICAL GENOMICS, 2001, 7 (02) :97-104
[5]
Ribosomal highly basic 23-kDa protein as a reliable standard for gene expression analysis [J].
Jesnowski, R ;
Backhaus, C ;
Ringel, J ;
Löhr, M .
PANCREATOLOGY, 2002, 2 (04) :421-424
[6]
Control genes and variability: Absence of ubiquitous reference transcripts in diverse mammalian expression studies [J].
Lee, PD ;
Sladek, R ;
Greenwood, CMT ;
Hudson, TJ .
GENOME RESEARCH, 2002, 12 (02) :292-297
[7]
Glyceraldehyde-3-phosphate dehydrogenase expression varies with age and nutrition status [J].
Mozdziak, PE ;
Dibner, JJ ;
McCoy, DW .
NUTRITION, 2003, 19 (05) :438-440
[8]
*NIH, 2005, CANC GEN AN PROJ
[9]
Validation of endogenous controls for gene expression analysis in microdissected human renal biopsies [J].
Schmid, H ;
Cohen, CD ;
Henger, A ;
Irrgang, S ;
Schlöndorff, D ;
Kretzler, M .
KIDNEY INTERNATIONAL, 2003, 64 (01) :356-360
[10]
Housekeeping genes as internal standards: use and limits [J].
Thellin, O ;
Zorzi, W ;
Lakaye, B ;
De Borman, B ;
Coumans, B ;
Hennen, G ;
Grisar, T ;
Igout, A ;
Heinen, E .
JOURNAL OF BIOTECHNOLOGY, 1999, 75 (2-3) :291-295