Ribosomal highly basic 23-kDa protein as a reliable standard for gene expression analysis.
OBJECTIVE
Analysis of gene expression is dependent on normalization using housekeeping genes. However, many of these housekeeping genes (e.g. GAPDH, beta-actin) are upregulated in chronic pancreatitis and pancreatic cancer, and cannot be used for normalization. For this reason we tried to identify a housekeeping gene useful for expression analysis in pancreatic diseases.


METHODS
RNA isolated from various tissues and states of disease was subjected to reverse transcription and subsequently amplified by PCR using primers for GAPDH and for the ribosomal highly basic 23-kDa (rb 23-kDa, RPL13A) protein.


RESULTS
As anticipated, expression of GAPDH varied markedly in the different tissues, whereas the expression of rb 23-kDa was constant in all samples investigated.


CONCLUSIONS
We recommend the use of the ribosomal highly basic 23-kDa protein as a standard for normalization at least for the pancreas and the prostate.
Department of Medicine II, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany. ralf.jesenofsky@med.ma.uni-heidelberg.de
R
R
Jesnowski
C
C
Backhaus
J
J
Ringel
M
M
Löhr
D015870:Gene Expression; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D006801:Humans; D008970:Molecular Weight; D010182:Pancreatic Diseases / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012269:Ribosomal Proteins / Q000737:chemistry* / Q000235:genetics*
D012333:RNA, Messenger; D012269:Ribosomal Proteins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases