Journal
JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS
Volume 70, Issue 3, Pages 481-486Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbbm.2006.11.008
Keywords
gene expression analysis; housekeeping gene; acidic ribosomal phosphoprotein P0 (36B4); northern analysis
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Housekeeping genes are often used as internal standards for gene expression analysis. When steady-state transcript levels of 4 typically used housekeeping genes, i.e., beta-actin, glyceraldehyde 3-phosphate dehydrogenase, cyclophilin, and acidic ribosomal phosphoprotein PO (304), were evaluated in various rat tissues, the 36B4 gene seemed to be the most suitable as a standard to compare the expression levels of genes among different tissues. Next, for possible quantitative comparison of the expression level of this gene among different animal species, we compared the nucleotide sequence of the cDNA of 36B4 among rats, mice, and humans. As a result, highly conserved regions showing more than 97.5% identities were observed in the 5' portion of its open reading frame. When samples of synthesized mRNA encoding rat, mouse, and human 36134 were hybridized with the entire cDNA encoding rat 36B4 as a probe, hybridization signals of mRNAs of mouse and human 36134 were much weaker than those of mRNA encoding rat 36B4. However, when they were hybridized with an oligonucleotide probe corresponding to the highly conserved regions, they showed similar signal intensities. Thus, these highly conserved regions of the cDNA encoding 36134 were concluded to be an effective standard for use in gene expression analysis. (c) 2006 Elsevier B.V. All rights reserved.
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