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
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- 作者：Rie Akamine ; Takenori Yamamoto ; Masahiro Watanabe ; Naoshi Yamazaki ; Masatoshi Kataoka ; Mitsuru Ishikawa ; Toshihiko Ooie ; Yoshinobu Baba ; Yasuo Shinohara
- 关键词：Gene expression analysis ; Housekeeping gene ; Acidic ribosomal phosphoprotein P0 (36B4) ; Northern analysis
- 刊名：Journal of Biochemical and Biophysical Methods
- 出版年：2007
- 出版时间：10 April 2007
- 年：2007
- 卷：70
- 期：3
- 页码：481-486
- 全文大小：752 K
文摘
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., β-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.