Detection of the human platelet-activating factor receptor mRNA in human B lymphocytes by polymerase chain reaction on reverse transcripts (RT-PCR)
The platelet activating factor receptor (hPAFR) is a GTP binding protein linked receptor of the rhodopsin family. The hPAFR gene maps to chromosome 1 and is characterized by the absence of introns in its coding region. Because PAF demonstrates unique properties as a growth factor in human B lymphoblastoid cell lines, we developed an RT-PCR in order to detect the presence of hPAFR mRNA. We examined the presence of the hPAFR in a series of B lymphoblastoid cell lines, as well as on fresh human B cells and the T-leukemia cell line Jurkat. All cells of B lineage expressed hPAFR mRNA, including a B cell line not previously shown to express functional hPAFR. In contrast, the T-leukemia cell line Jurkat expressed very low levels of hPAFR mRNA. We discuss the methodology and its validation in developing an RT-PCR for intronless genes such as the hPAFR.
Bastien
Y
Y
McGill University/Montreal Children's Hospital Research Institute, Quebec, Canada.
Mazer
B D
BD
D001402:B-Lymphocytes / Q000378:metabolism*; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D006801:Humans; D008969:Molecular Sequence Data; D010972:Platelet Activating Factor / Q000378:metabolism; D010980:Platelet Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D010972:Platelet Activating Factor; D010980:Platelet Membrane Glycoproteins; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; C039351:platelet activating factor receptor