【摘要】：應用RT-PCR技術,從小鼠脾淋巴細胞中克隆AIM 2基因,通過生物信息學軟件對AIM2的遺傳演化關系、分子結構特征以及氨基酸序列進行分析,并應用q RT-PCR檢測在不同組織中的m RN A轉錄水平。結果顯示,克隆的小鼠AIM2基因開放閱讀框全長1 065 bp,編碼354個氨基酸,與其他動物的AIM2基因序列具有較高的同源性(56.9%~88.9%);q RT-PCR檢測表明,AIM2基因在肌肉和心臟組織中轉錄水平較高。此外,將AIM2基因亞克隆至真核表達載體p CMV-Tag 2B進行W estern-blot驗證,然后將正確的重組表達質粒轉染H EK 293T細胞,進而應用雙熒光素酶報告系統(tǒng)檢測證實在poly(d A-d T)和poly(d G-d C)刺激下,過表達小鼠AIM2對轉錄因子NF-κB和AP-1的活化過程有顯著增強作用。上述研究結果表明,小鼠AIM2能夠識別富含A/T或C/G的D NA,是重要的胞漿DNA識別受體,這為進一步探討AIM2與DNA病毒的相互作用機制奠定基礎。
[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.
【作者單位】： 西北民族大學生命科學與工程學院;中國農業(yè)科學院蘭州獸醫(yī)研究所家畜疫病病原生物學重點實驗室農業(yè)部獸醫(yī)公共衛(wèi)生重點實驗室;甘肅農業(yè)大學動物醫(yī)學院;
【基金】：國家自然科學基金項目(31302072;31372423;30871884)
【分類號】：S852.42

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1 劉興樓;固有免疫細胞AIM2炎性體通路在抗HCMV免疫機制中的作用[D];華中科技大學;2011年

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