Article

Selection of reference genes for gene expression studies in human neutrophils by real-time PCR

BMC Molecular Biology 01/2005;
Source: DOAJ

ABSTRACT Abstract

Background

Reference genes, which are often referred to housekeeping genes, are frequently used to normalize mRNA levels between different samples. However the expression level of these genes may vary among tissues or cells, and may change under certain circumstances. Thus the selection of reference gene(s) is critical for gene expression studies. For this purpose, 10 commonly used housekeeping genes were investigated in isolated human neutrophils.

Results

Initial screening of the expression pattern demonstrated that 3 of the 10 genes were expressed at very low levels in neutrophils and were excluded from further analysis. The range of expression stability of the other 7 genes was (from most stable to least stable): GNB2L1 (Guanine nucleotide binding protein, beta polypeptide 2-like 1), HPRT1 (Hypoxanthine phosphoribosyl transferase 1), RPL32 (ribosomal protein L32), ACTB (beta-actin), B2M (beta-2-microglobulin), GAPD (glyceraldehyde-3-phosphate dehydrogenase) and TBP (TATA-binding protein). Relative expression levels of the genes (from high to low) were: B2M, ACTB, GAPD, RPL32, GNB2L1, TBP, and HPRT1.

Conclusion

Our data suggest that GNB2L1, HPRT1, RPL32, ACTB, and B2M may be suitable reference genes in gene expression studies of neutrophils.

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Keywords

7 genes
 
ACTB
 
beta polypeptide 2-like 1
 
different samples
 
expression level
 
expression pattern
 
expression stability
 
GAPD
 
gene expression studies
 
glyceraldehyde-3-phosphate dehydrogenase
 
Guanine nucleotide binding protein
 
HPRT1
 
Hypoxanthine phosphoribosyl transferase 1
 
low levels
 
normalize mRNA levels
 
reference gene(s)
 
Relative expression levels
 
ribosomal protein L32
 
stable
 
TATA-binding protein
 

Xiaozhu Zhang