The New Favorite Reference Gene: 36B4, a.k.a. RPLP0

Posted on Wed, Oct 14, 2009 @ 08:06 AM

A "Referral from the Doctor" Blog Article-

Ribosomes are comprised of two subunits, one small (40S) and one large (60S). The 36B4 gene encodes an acidic ribosomal phosphoprotein P0 (RPLP0), which forms tight associations with the smaller 40S protein (L12). 36B4 protein is part of a pentameric complex that forms a stem-like structure, protruding into the cytoplasm, off of the 60S subunit. This protrusion functions to support the GTPase steps in the translocation of protein synthesis. Why is 36B4 fast becoming the new reference gene for probe-based real time RT-PCR?

The 36B4 cDNA nucleotide sequence has highly conserved regions in the 5-prime end of its open reading frame that cross tissue and species boundaries. When compared to the transcript levels of other common reference genes, such as beta-actin and cyclophilin, 36B4 proved to be a very reliable and consistent standard for use in gene expression analysis among multiple tissues including: brain, heart, liver, kidney, muscle and lung. Furthermore, the nucleotide sequence homology between species is remarkably high. For canis familiaris (dog) the homology to the human sequence is 94%; for pan troglodytes (chimpanzee), 99%; and for bos taurus (bovine), 94%. In basic science the most commonly used species are mus musculus (mouse) and rattus norvegicus (rat) whose 36B4 maintains a homology with the human gene of 88% and 89% respectively. As with other highly expressed genes, 36B4 has 8 related pseudo-genes in the human genome, found on chromosomes 1-3, 11, 14, 15 and 18.

Recent trends have supported the use of multiple reference genes as normalizing factors in real-time RT-PCR. For a list of the most highly recommended reference genes, see our blog article, What Genes are Hot, What Genes are Not.

Written by: Christina Ferrell, Ph.D., Technical Applications Specialist

References:

Rie Akamine , T. Yamamoto, M. Watanabe, N. Yamazaki, M. Kataoka, M. Ishikawa, T. Ooie, Y. Baba, Y. Shinohara. 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. (2007) J. Biochem. Biophys. Methods 70: 481-486

Jorge Laborda. 36B4 cDNA used as an estradiol-independent mRNA control is the cDNA for human acidic ribosomal phosphoprotein P0. (1991) Nucleic Acids Research 19 (14): 3998
Key words: 36B4, RPLP0, acidic ribosomal phosphoprotein P0

Topics: Real-time PCR, Custom Oligos, Referral from the Doctor

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