不同品种羊血液和肺脏中TLRs基因转录水平差异分析

Differential analysis of transcript levels of TLRs genes in the blood and lungs of different breeds of sheep

【目的】分析不同品种羊Toll样受体(TLRs)基因转录水平的差异性,为揭示TLRs基因转录水平与羊疫病间的关联性提供参考依据。【方法】选取贵州省主要饲养的贵州黑山羊、贵州白山羊、黔北麻羊、波尔山羊和湖羊为研究对象,根据GenBank已公布的TLRs基因序列设计荧光定量PCR特异性扩增引物及TaqMan探针引物,利用TaqMan探针荧光定量PCR检测不同品种羊血液和肺脏中TLR1~TLR10基因转录水平差异。【结果】TLR1~TLR10基因在不同品种羊血液和肺脏中均有转录表达。不同品种羊血液和肺脏中的TLRs基因转录水平均存在差异性,在波尔山羊血液中TLR2、TLR4和TLR5基因转录水平均极显著低于其他4种羊(P<0.01,下同),TLR7和TLR8基因转录水平则极显著低于除黑山羊外的其他3种羊;在白山羊血液中TLR4、TLR7、TLR8和TLR9基因转录水平极显著高于其他4种羊;在湖羊肺脏中TLR2、TLR3、TLR4、TLR5和TLR8基因转录水平极显著低于其他4种羊,而在黔北麻羊肺脏中TLR5和TLR8基因转录水平极显著高于其他4种羊。此外,在5种羊血液中TLR2、TLR4、TLR7和TLR8基因转录水平均极显著高于其他TLRs基因转录水平;羊肺脏中TLR2、TLR3、TLR4、TLR5和TLR8基因转录水平相对较高,在贵州白山羊、贵州黑山羊和波尔山羊均表现为极显著高于其他TLRs基因转录水平。【结论】不同品种羊血液和肺脏中TLRs基因转录水平具有差异性,以TLR2、TLR3、TLR4、TLR5和TLR8基因转录水平相对较高,提示TLRs在不同品种羊机体中发挥着清除病原体及维持机体稳态的作用,而TLRs基因转录水平差异可能是造成不同品种羊对疫病易感差异的原因之一。

【Objective】To analyze the variability of transcript levels of Toll-like receptors(TLRs)genes in different breeds of sheep,and to provide a reference basis for revealing the association between the transcript levels of TLRs genes and epidemic diseases in sheep.【Method】In this study,Guizhou black goat,Guizhou white goat,Qianbei Ma goat,Boer goat and Hu sheep,which are the main breeding species in Guizhou Province,were selected for the study,and fluores-cent quantitative PCR specific amplification primers and TaqMan probe primers were designed according to the published TLRs gene sequences in GenBank,and the differences in transcript levels of TLR1-TLR10 genes in blood and lung of dif-ferent breeds of sheep were detected by using TaqMan probe fluorescent quantitative PCR.【Result】TLR1-TLR10 genes were transcriptionally expressed in the blood and lungs of different breeds of sheep.Differential transcript levels of TLRs genes were in both blood and lung of different breeds of sheep,the transcript levels of TLR2,TLR4 and TLR5 genes in the blood of Boer goats were extremely significantly lower than those of the other four species(P<0.01,the same below),the transcript levels of TLR7 and TLR8 genes were extremely significantly lower than those of the other three species of sheep except black goats.The transcript levels of TLR4,TLR7,TLR8 and TLR9 genes in the blood of white goats were extreme-ly significantly higher than those of the other four species.The transcript levels of TLR2,TLR3,TLR4,TLR5 and TLR8 genes in the lungs of Hu sheep were extremely significantly lower than those of the other four sheep species,while the transcript levels of TLR5 and TLR8 genes were extremely highly significant higher in the lungs of Qianbei Ma sheep than in the other four species.In addition,the transcript levels of TLR2,TLR4,TLR7 and TLR8 genes in the blood of five sheep species were all extremely significantly higher than the transcript levels of other TLRs genes.Transcript levels of TLR2,TLR3,TLR4,TLR5 and TLR8 genes in sheep lungs were relatively high,in Guizhou white goat,Guizhou black goat and Boer goat all showed extremely significantly higher transcript levels than other TLRs genes.【Conclusion】The transcript levels of TLRs genes in the blood and lungs of different breeds of sheep are different,with TLR2,TLR3,TLR4,TLR5 and TLR8 genes having relatively high transcript levels.It is suggested that TLRs play a role in the organism of different breeds of sheep in scavenging pathogens and maintaining the homeostasis of the organism,and the difference in the transcription level of TLRs genes may be one of the reasons for the difference in susceptibility of different breeds of sheep to epidemic diseases.