Detection of human interferon-alpha-encoding gene expression.
We have devised a sensitive method based on the polymerase chain reaction (PCR) to detect expression of human interferon-alpha-encoding genes (IFN-A) in general, and specifically, expression of the IFN-A2 or IFN-A4 genes. The utility of the PCR approach was assessed by analysis of cloned IFN-A genes, as well as genomic DNA and mRNA isolated from peripheral blood mononuclear cells. We demonstrate the specific amplification of sequences encoding IFN subtypes IFN-alpha-2 and IFN-alpha-4 from as little as 0.1 pg of IFN-A mRNA. The PCR technique has potential clinical application for the detection of IFN-A expression and, thus, identification of the IFN-alpha subtypes produced, particularly in small biopsy samples or otherwise, where only low numbers of cells are available.
Centre for Molecular Biology and Medicine, Monash University, Clayton, Victoria, Australia.
E R
ER
Brandt
R J
RJ
Devenish
B F
BF
Cheetham
A W
AW
Linnane
D001483:Base Sequence; D015139:Blotting, Southern; D002899:Chromosomes, Human, Pair 9; D003001:Cloning, Molecular / Q000379:methods; D004247:DNA / Q000097:blood / Q000235:genetics / Q000302:isolation & purification; D015870:Gene Expression; D006801:Humans; D016898:Interferon-alpha / Q000235:genetics*; D008969:Molecular Sequence Data; D009000:Monocytes / Q000502:physiology; D009838:Oligodeoxyribonucleotides; D010222:Parainfluenza Virus 1, Human / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000097:blood / Q000235:genetics / Q000302:isolation & purification; D014407:Tumor Cells, Cultured
D016898:Interferon-alpha; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D004247:DNA