Detection of bovine interleukin 1 alpha and interleukin 1 beta gene expression by reverse transcription-polymerase chain reaction.
Interleukin 1 (IL-1) is a cytokine that mediates a variety of immunoregulatory and inflammatory activities. Its role in proliferation of helper T cells is particularly important in the initiation of immune responses. Bovine interleukin 1 alpha (IL-1 alpha) and interleukin 1 beta (IL-1 beta) mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR) technique in bovine peripheral blood mononuclear cells (PBMCs). Poly (A) + RNAs isolated from PBMCs by guanidium thiocyanate/oligo (dT)-cellulose chromatography extraction, were reverse-transcribed and the complementary DNAs amplified in a polymerase chain reaction primed with bovine IL-1 alpha and IL-1 beta sequence-specific primers. RT-PCR products were analyzed by agarose gel electrophoresis. As a result of the amplification of specific cDNAs by 35 cycles of PCR, the IL-1 alpha mRNA was detected from 0.01 ng of poly (A) + RNA, and IL-1 beta mRNA was detected from 0.1 pg of poly (A) + RNA isolated from PBMCs stimulated with lipopolysaccharide (LPS) for 4 h. Using 5 ng of poly (A) + RNA isolated from fresh bovine PBMCs unstimulated with any mitogen as template for RT-PCR, both IL-1 alpha and IL-1 beta mRNAs were detected at a low level.
Ito
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T
Hokkaido Branch, National Institute of Animal Health, Sapporo, Japan.
Kodama
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M
D000818:Animals; D001483:Base Sequence; D002417:Cattle / Q000276:immunology*; D017931:DNA Primers; D015870:Gene Expression; D007375:Interleukin-1 / Q000235:genetics / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D012333:RNA, Messenger / Q000032:analysis*; D014158:Transcription, Genetic
D017931:DNA Primers; D007375:Interleukin-1; D012333:RNA, Messenger