Usefulness of the 5' region of the cDNA encoding acidic ribosomal phosphoprotein P0 conserved among rats, mice, and humans as a standard probe for gene expression analysis in different tissues and animal species.
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 P0 (36B4), 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 36B4 were hybridized with the entire cDNA encoding rat 36B4 as a probe, hybridization signals of mRNAs of mouse and human 36B4 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 36B4 were concluded to be an effective standard for use in gene expression analysis.
Akamine
Rie
R
Institute for Genome Research, Faculty of Pharmaceutical Sciences, University of Tokushima, Kuramotocho-3, Tokushima 770-8503, Japan.
Yamamoto
Takenori
T
Watanabe
Masahiro
M
Yamazaki
Naoshi
N
Kataoka
Masatoshi
M
Ishikawa
Mitsuru
M
Ooie
Toshihiko
T
Baba
Yoshinobu
Y
Shinohara
Yasuo
Y
D000818:Animals; D001483:Base Sequence; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000235:genetics*; D020869:Gene Expression Profiling; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D012269:Ribosomal Proteins / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D014018:Tissue Distribution; D014158:Transcription, Genetic
D018076:DNA, Complementary; D010750:Phosphoproteins; D012333:RNA, Messenger; D012269:Ribosomal Proteins; C054675:ribosomal protein P0