SPLUNC1基因在2种绵羊不同组织器官中差异表达分析  

Differential expression of the SPLUNC1 gene in different tissues and organs of two breeds of sheep

作　　者：李珂儿 李劼[1] 李增强 魏立翔 孙延鸣[1] LI Ke'er;LI Jie;LI Zengqiang;WEI Lixiang;SUN Yanming(College of Animal Science and Technology,Shihezi University,Shihezi 832000,China)

机构地区：[1]石河子大学动物科技学院,新疆石河子832000

出　　处：《畜牧与兽医》2021年第7期89-94,共6页Animal Husbandry & Veterinary Medicine

基　　金：兵团重大科技项目(2017AA006)。

摘　　要：SPLUNC1是抗病原微生物感染的关键蛋白。为研究SPLUNC1基因在2种绵羊不同组织器官中的差异表达情况,试验选取健康新疆阿勒泰羊5只,多胎萨福克羊5只,放血处死后分别采集气管、支气管、肺脏、胃、食管、结肠、心脏、脾脏、皮肤,提取RNA,反转录合成cDNA。以cDNA为模板,扩增SPLUNC1基因748 bp特征片段,克隆至pMD19-T载体中,提取质粒。以梯度稀释的质粒为模板,建立SPLUNC1基因转录本拷贝数的绝对定量标准曲线,对2种绵羊各组织器官中SPLUNC1的mRNA初始拷贝数进行定量分析。结果表明:阿勒泰羊标准曲线扩增效率(E)为97%,回归系数(R^(2))为0.9990,SPLUNC1 mRNA初始拷贝数从高至低依次为气管、支气管、肺脏、脾脏、食管、心脏、胃、结肠、皮肤。萨福克羊标准曲线扩增效率(E)为99%,回归系数(R^(2))为0.9989,SPLUNC1 mRNA初始拷贝数从高至低依次为气管、支气管、肺脏、脾脏、心脏、食管、胃、结肠、皮肤。2种绵羊的气管SPLUNC1 mRNA表达量均显著高于其他各组织(P<0.05),皮肤表达量最低。新疆阿勒泰羊的9种组织器官该基因表达量均显著高于多胎萨福克羊(P<0.05)。上述结果说明绵羊的SPLUNC1基因的分布具有组织器官特异性,为揭示绵羊抗感染的分子调控机制提供一定参考。SPLUNC1 is a key protein in resistance to pathogenic microorganism infection.In order to study the differential expression of the SPLUNC1 gene in different tissues and organs of two breeds of sheep,5 healthy Xinjiang Altay sheep and 5 multiparous Suffolk sheep were used here.After bloodletting slaughter of the sheep,the samples of trachea,bronchus,lung,stomach,esophagus,colon,heart,spleen and skin were collected from the animals to extract RNA and to perform reverse transcription to synthesize cDNA.A 748 BP characteristic fragment of the SPLUNC1 gene was amplified from cDNA and cloned into a pmd19-t vector.Then,gradiently diluted plasmids were used as template to establish an absolute quantitative standard curve of the SPLUNC1 gene transcript copy number,and to quantitatively analyzed the initial copy number of SPLUNC1 mRNA in different tissues and organs of the two breeds of sheep.The results showed that the amplification efficiency(E)of the standard curve of the Altay sheep was 97%,and the regression coefficient(R^(2))was 0.9990.The initial copy numbers of SPLUNC1 mRNA from high to low in their organs were in the order of the trachea,bronchus,lung,spleen,esophagus,heart,stomach,colon and skin.The standard curve(E)of the Suffolk sheep was 99%,and the regression coefficient(R^(2))was 0.9989.The initial copy numbers of SPLUNC1 mRNA from high to low in their organs were in the order of the trachea,bronchus,lung,spleen,heart,esophagus,stomach,colon and skin.The expression of SPLUNC1 mRNA in the trachea of the breeds of sheep was significantly higher than that in the other tissues(P<0.05),with the lowest expression in the skin.These results indicated that the distribution of the SPLUNC1 gene in sheep was tissue-and organ-specific,which provides reference for research on the molecular regulation mechanism of anti-infection in sheep.

关 键 词：多胎萨福克羊 阿勒泰羊 SPLUNC1基因 实时定量PCR 组织表达 

分 类 号：S852.23[农业科学—基础兽医学]