小鼠AIM2分子结构特征及天然免疫模式识别功能的研究
发布时间：2018-11-12 18:32
【摘要】：应用RT-PCR技术,从小鼠脾淋巴细胞中克隆AIM 2基因,通过生物信息学软件对AIM2的遗传演化关系、分子结构特征以及氨基酸序列进行分析,并应用q RT-PCR检测在不同组织中的m RN A转录水平。结果显示,克隆的小鼠AIM2基因开放阅读框全长1 065 bp,编码354个氨基酸,与其他动物的AIM2基因序列具有较高的同源性(56.9%~88.9%);q RT-PCR检测表明,AIM2基因在肌肉和心脏组织中转录水平较高。此外,将AIM2基因亚克隆至真核表达载体p CMV-Tag 2B进行W estern-blot验证,然后将正确的重组表达质粒转染H EK 293T细胞,进而应用双荧光素酶报告系统检测证实在poly(d A-d T)和poly(d G-d C)刺激下,过表达小鼠AIM2对转录因子NF-κB和AP-1的活化过程有显著增强作用。上述研究结果表明,小鼠AIM2能够识别富含A/T或C/G的D NA,是重要的胞浆DNA识别受体,这为进一步探讨AIM2与DNA病毒的相互作用机制奠定基础。
[Abstract]:The AIM 2 gene was cloned from mouse spleen lymphocytes by RT-PCR technique. The genetic evolution, molecular structure and amino acid sequence of AIM2 were analyzed by bioinformatics software. Q RT-PCR was used to detect the m RN A transcription level in different tissues. The results showed that the total length of mouse AIM2 gene was 1 065 bp, encoding 354 amino acids, which had high homology with other animal AIM2 gene sequences (56.9% or 88.9%). Q RT-PCR analysis showed that the transcription level of AIM2 gene was higher in muscle and heart tissues. In addition, the AIM2 gene was subcloned into eukaryotic expression vector p CMV-Tag 2B for W estern-blot validation, and then the correct recombinant expression plasmid was transfected into H EK 293T cells. Furthermore, double luciferase reporting system was used to detect the activation process of NF- 魏 B and AP-1 induced by poly (d A-d T) and poly (d G-d C). These results suggest that mouse AIM2 can recognize D NA, which is rich in A / T or C / G, as an important cytoplasmic DNA recognition receptor, which lays a foundation for further study of the interaction mechanism between AIM2 and DNA virus.
【作者单位】： 西北民族大学生命科学与工程学院;中国农业科学院兰州兽医研究所家畜疫病病原生物学重点实验室农业部兽医公共卫生重点实验室;甘肃农业大学动物医学院;
【基金】：国家自然科学基金项目(31302072;31372423;30871884)
【分类号】：S852.42
本文编号：2327877
[Abstract]:The AIM 2 gene was cloned from mouse spleen lymphocytes by RT-PCR technique. The genetic evolution, molecular structure and amino acid sequence of AIM2 were analyzed by bioinformatics software. Q RT-PCR was used to detect the m RN A transcription level in different tissues. The results showed that the total length of mouse AIM2 gene was 1 065 bp, encoding 354 amino acids, which had high homology with other animal AIM2 gene sequences (56.9% or 88.9%). Q RT-PCR analysis showed that the transcription level of AIM2 gene was higher in muscle and heart tissues. In addition, the AIM2 gene was subcloned into eukaryotic expression vector p CMV-Tag 2B for W estern-blot validation, and then the correct recombinant expression plasmid was transfected into H EK 293T cells. Furthermore, double luciferase reporting system was used to detect the activation process of NF- 魏 B and AP-1 induced by poly (d A-d T) and poly (d G-d C). These results suggest that mouse AIM2 can recognize D NA, which is rich in A / T or C / G, as an important cytoplasmic DNA recognition receptor, which lays a foundation for further study of the interaction mechanism between AIM2 and DNA virus.
【作者单位】： 西北民族大学生命科学与工程学院;中国农业科学院兰州兽医研究所家畜疫病病原生物学重点实验室农业部兽医公共卫生重点实验室;甘肃农业大学动物医学院;
【基金】：国家自然科学基金项目(31302072;31372423;30871884)
【分类号】：S852.42
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1 刘兴楼;固有免疫细胞AIM2炎性体通路在抗HCMV免疫机制中的作用[D];华中科技大学;2011年
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