【摘要】：泛素連接酶E3家族在脊椎動物的先天免疫中發(fā)揮重要作用,由于RING finger結構域的存在,三重基序(tripartite motif,TRIM)蛋白家族的很多成員被報道具有泛素連接酶活性。為了研究TRIM16和TRIM25基因在尼羅羅非魚(Oreochromis niloticus)免疫過程中的作用,本研究通過逆轉錄PCR和RACE獲得了尼羅羅非魚TRIM16和TRIM25基因的cDNA及基因組序列,并進行了基因結構和蛋白二級結構分析,qRT-PCR檢測基因的組織分布、在胚胎發(fā)育過程中表達變化及其對無乳鏈球菌(Streptococcus agalactiae)感染的響應。結果表明,TRIM16基因的cDNA(GenBank登錄號:KY746714)全長2 314 bp,ORF 1 677 bp,編碼558個氨基酸;TRIM25基因的cDNA(GenBank登錄號:KY968697)全長2 748 bp,ORF1 677 bp,編碼558個氨基酸,二者基因組序列均無內(nèi)含子。氨基酸序列分析顯示,TRIM16和TRIM25均具有TRIM家族的RING finger結構域、B-box結構域等保守結構。組織分析表明,在所檢測的11個組織和器官中,TRIM16和TRIM25基因均有表達,血液中表達最高,肝臟中表達最低,血液中的表達量分別為肝臟(對照組)的60.46和274.07倍。這兩個基因在尼羅羅非魚胚胎發(fā)育過程中均有表達。人工感染無乳鏈球菌后,TRIM16和TRIM25基因在所檢測的5個組織和器官中的表達量均存在升高的階段,TRIM16基因在腸、脾、鰓、腎臟和血液中的最高表達量分別為0 h(對照組)的1.30、2.09、1.61、7.81和6.05倍,TRIM25基因在腸、脾、鰓、腎臟和血液中的最高表達量分別為對照組的11.13、1.22、1.26、61.41和77.80倍。上述結果表明,TRIM16和TRIM25基因在尼羅羅非魚抗無乳鏈球菌感染的過程中發(fā)揮了重要作用。研究結果為進一步了解羅非魚的抗感染免疫機制、探索病害防治新途徑提供了理論基礎。
[Abstract]:Ubiquitin ligase E3 family plays an important role in innate immunity of vertebrates. Due to the existence of RING finger domain, many members of the triple motif (tripartite motif,TRIM) protein family have been reported to have ubiquitin ligase activity. In order to study the role of TRIM16 and TRIM25 genes in (Oreochromis niloticus) immunization of Nile tilapia, the cDNA and genomic sequences of TRIM16 and TRIM25 genes of Nile tilapia were obtained by reverse transcription PCR and RACE. The gene structure and protein secondary structure were analyzed. QRT-PCR was used to detect the tissue distribution of the genes, the expression changes during embryonic development and their response to Streptococcus lactis (Streptococcus agalactiae) infection. The results showed that the full length of TRIM16 gene was 2314 bp,ORF1 677 bp, encoding 558 amino acids, and the full length of TRIM25 gene was 2748 bp,ORF1 677 bp, encoding 558 amino acids, both of which had no introns. Amino acid sequence analysis showed that both TRIM16 and TRIM25 had conserved structures such as RING finger domain and B-box domain of TRIM family. Tissue analysis showed that TRIM16 and TRIM25 genes were expressed in 11 tissues and organs, the highest expression was found in blood, and the lowest expression was found in liver. The expression level in blood was 60.46 and 274.07 times higher than that in liver (control group), respectively. These two genes were expressed during embryonic development of Nile tilapia. After artificial infection with Streptococcus lactis, the expression of TRIM16 and TRIM25 genes in the five tissues and organs detected was increased. The highest expression of TRIM16 gene in intestine, spleen, Gill, kidney and blood was 1.30, 2.09, 1.61, 7.81 and 6.05 times of that in intestine, spleen, Gill, kidney and blood, respectively. TRIM25 gene was in intestine, spleen and Gill. The highest expression levels in kidney and blood were 11.13, 1.22, 1.26, 61.41 and 77.80 times higher than those in control group, respectively. These results suggest that TRIM16 and TRIM25 genes play an important role in the prevention of Streptococcus lactis infection in Nile tilapia. The results provide a theoretical basis for further understanding the anti-infection immune mechanism of tilapia and exploring a new way of disease control.
【作者單位】： 中國水產(chǎn)科學研究院珠江水產(chǎn)研究所/農(nóng)業(yè)部熱帶亞熱帶水產(chǎn)資源利用與養(yǎng)殖重點實驗室;上海海洋大學水產(chǎn)與生命學院;
【基金】：現(xiàn)代農(nóng)業(yè)產(chǎn)業(yè)技術體系專項(CARS-49) 廣東省創(chuàng)新載體建設項目(No.2013B090700004) 廣州市科技計劃項目產(chǎn)學研協(xié)同創(chuàng)新重大專項(No.201508020046)
【分類號】：S917.4

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