Analysis of interleukin receptor gene expression in mouse fetal liver by quantitative polymerase chain reaction.
We describe a simplified and sensitive polymerase chain reaction (PCR)-based method for the quantification of low-abundance RNA for mouse cytokine receptor genes. Accurate quantification is achieved in a two-step protocol which uses a synthetic RNA as an internal standard. The proper titration of the amount of mRNA molecules is followed by a kinetic analysis which ensures precise measurement. This quantitative PCR method provides a rapid and reliable way to quantify the amount of cytokine receptor mRNA in samples containing as few as 1000 molecules of RNA for a cytokine receptor target gene. We illustrate our approach by quantifying mRNA levels for two families of cytokine receptor genes in the fetal liver and bone marrow of the mouse. Our results reveal early and abundant expression of the genes encoding the signal transducing subunits interleukin-2 receptor gamma and gp130. Their expression seems to precede that of the genes encoding the specific subunits of these interleukin receptor systems.
Jia
G Q
GQ
Center for Blood Research, Inc., Harvard Medical School, Boston, MA 02115, USA.
Gutierrez-Ramos
J C
JC
D000818:Animals; D015703:Antigens, CD; D001483:Base Sequence; D050822:Cytokine Receptor gp130; D017931:DNA Primers / Q000737:chemistry; D015870:Gene Expression; D008099:Liver / Q000196:embryology* / Q000276:immunology; D008562:Membrane Glycoproteins / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000235:genetics; D018123:Receptors, Interleukin / Q000235:genetics*; D015375:Receptors, Interleukin-2 / Q000235:genetics
D015703:Antigens, CD; D017931:DNA Primers; C494200:Il6st protein, mouse; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D018123:Receptors, Interleukin; D015375:Receptors, Interleukin-2; D050822:Cytokine Receptor gp130