[1]杨 源,王 军,唐 沙,等.TRAF6、AP-1、NF-кB基因TaqMan探针荧光定量检测方法的建立及应用[J].中国预防兽医学报,2019,(03):261-266.[doi:0.3969/j.issn.1008-0589.201807022]
 YANG Yuan,WANG Jun,TANG Sha,et al.Establishment and application of real-time PCR methods for TaqMan probes of the Toll-like receptor MyD88 signaling pathway linkers in goats[J].Chinese journal of preventive veterinary medicine,2019,(03):261-266.[doi:0.3969/j.issn.1008-0589.201807022]
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TRAF6、AP-1、NF-кB基因TaqMan探针荧光定量检测方法的建立及应用()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2019年03
页码:
261-266
栏目:
诊断和检测技术
出版日期:
2019-04-25

文章信息/Info

Title:
Establishment and application of real-time PCR methods for TaqMan probes of the Toll-like receptor MyD88 signaling pathway linkers in goats
文章编号:
1008-0589(2019)03-0261-06
作者:
 

杨 源12王 军12唐 沙12岳 筠3周 怡12文 明12周碧君12程振涛12*

 (1. 贵州大学 动物科学学院,贵州 贵阳 550025;2. 贵州省动物疫病与兽医公共卫生重点实验室,贵州 贵阳 550025;
3. 贵州省动物疫病预防控制中心,贵州 贵阳 550008)
Author(s):
YANG Yuan12 WANG Jun12 TANG Sha12 YUE Jun3 ZHOU Yi12 WEN Ming12ZHOU Bi-jun12 CHENG Zhen-tao12*
 (1. College of Animal Science, Guizhou University, Guiyang 550025, China; 2. Key Laboratory of Animal Diseases and
Veterinary Public Health in Guizhou Province Guiyang 550025, China; 3. Animal Disease Prevention and Control Center
in Guizhou Province, Guiyang 550008,China)
关键词:
Toll样受体MyD88信号通路分子
Keywords:
sheep Toll-like receptor MyD88 signaling pathway linker
分类号:
S852.6
DOI:
0.3969/j.issn.1008-0589.201807022
文献标志码:
A
摘要:
为探究不同品种羊Toll样受体MyD88通路重要接头分子在其肺组织中转录水平的差异,本研究建立了可用于分析不同品种羊Toll样受体MyD88信号通路接头分子MyD88、AP-1、NF-кB、TRAF6基因的TaqMan荧光定量PCR方法,并对该方法进行特异性、敏感性、重复性及应用性等性能评价。结果显示:建立的Toll样受体MyD88通路接头分子4个基因的TaqMan探针荧光定量PCR方法具有较强特异性和较高灵敏度,组内重复和组间重复的变异系数均在5 %以下,扩增效率均在90 %~105 %。将该方法初步应用于临床组织样本的检测,结果显示:NF-кB在黑山羊肺组织中转录水平极显著高于其余4种羊(p<0.01);在白山羊、麻羊、湖羊和波尔山羊肺组织中MyD88和TRAF6的转录水平极显著高于AP-1和NF-кB基因的转录水平(p<0.01)。本研究建立的方法为研究羊呼吸系统性疫病的发生与Toll样受体信号通路传导的关系奠定基础。
  
Abstract:
 

To explore the differences in the transcriptional levels of important linker molecules of the Toll-like receptor MyD88 pathway in lung tissue of different breeds of sheep, the real-time PCR methods were established based on the specific TaqMan probes for detecting the transcriptional levels of MyD88, AP-1, NF-кB and TRAF6 genes of the Toll-like receptor MyD88 signaling pathway linkers in different breeds of sheep. The results showed that the TaqMan PCR methods were high specificity and sensitivity, and the coefficient of variation of intra-group and inter-group assays were both below 5%, the amplification efficiency were between 90% and 105%. The transcription level changes of the linker molecules in the signal pathway could be quantitatively analyzed by the established standard curve. The methods were initially applied to the detection of clinical tissue samples and the results showed that NF-кB transcriptional level was significantly higher in black goat lung tissue than that in the other four goats (p<0.01). The mRNA levels of MyD88 and TRAF6 in the lung tissue of white goats, Ma sheep, Hu sheep and Boer goats were extremely higher than that of AP-1 and NF-кB mRNA levels (p<0.01). The established methods in this study lay a foundation for studying the relationship between the occurrence of respiratory disease in goats and the signal transduction of Toll-like receptor signaling pathway.

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                                (本文编辑:李 娜)

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更新日期/Last Update: 2019-04-26