Journal of Veterinary Medical Science
Online ISSN : 1347-7439
Print ISSN : 0916-7250
ISSN-L : 0916-7250
NOTE
Quantification of Llama Inflammatory Cytokine mRNAs by Real-Time RT-PCR
Raadan ODBILEGSatoru KONNAITatsufumi USUIKazuhiko OHASHIMisao ONUMA
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Keywords: Llama, LPS, real-time RT-PCR
JOURNAL FREE ACCESS

2005 Volume 67 Issue 2 Pages 195-198

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Abstract

We have developed a method by which llama cytokine mRNAs can be quantified using real-time reverse transcription polymerase chain reaction (RT-PCR). Total RNA was extracted from lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (PBMCs) of llama, reverse transcribed to cDNA, and cytokine profiles for interleukin (IL)-1α, IL-1β, IL-6 and tumor necrosis factor (TNF) α were quantified by real-time PCR. The expressions of mRNAs of inflammatory cytokines IL-1α, IL-1β, IL-6 and TNFα were upregulated upon stimulation with LPS in a dose- and time-dependent manner. Incubation of PBMCs with 100 and 1,000 pg/ml of LPS for 3 to 6 hr resulted in the acceleration of the mRNA levels of inflammatory cytokines. Here, we describe a highly sensitive and reproducible method to quantify the transcription of llama cytokine mRNAs by real-time RT-PCR with the double-stranded DNA-binding dye SYBR Green I.

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© 2005 by the Japanese Society of Veterinary Science

この記事はクリエイティブ・コモンズ [表示 - 非営利 - 改変禁止 4.0 国際]ライセンスの下に提供されています。
https://creativecommons.org/licenses/by-nc-nd/4.0/deed.ja
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