Quantification of Llama Inflammatory Cytokine mRNAs by Real-Time RT-PCR

Journal of Veterinary Medical Science

Raadan ODBILEG¹  Kazuhiko OHASHI¹  Satoru KONNAI¹  Misao ONUMA¹  Tatsufumi USUI¹ 

[1] Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University

关键词: Llama;    LPS;    real-time RT-PCR;   

DOI  :  10.1292/jvms.67.195

学科分类：兽医学

来源: Japanese Society of Veterinary Science

【 摘 要 】

References(17)Cited-By(7)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.

【 授权许可】

Unknown