【摘要】：為了研究含Armadillo重復(fù)蛋白8在凡納濱對(duì)蝦抗病感染過(guò)程中所起的免疫作用,本實(shí)驗(yàn)采用RACE-PCR技術(shù)首次克隆得到凡納濱對(duì)蝦ARMC8基因(LvARMC8,Gen Bank注冊(cè)號(hào):KX058562)的c DNA序列全長(zhǎng),并利用在線(xiàn)軟件進(jìn)行生物信息學(xué)分析;采用實(shí)時(shí)熒光定量PCR(Rt-PCR)技術(shù)對(duì)LvARMC8基因在凡納濱對(duì)蝦不同組織及感染白斑綜合征病毒和副溶血弧菌過(guò)程中的表達(dá)變化特征進(jìn)行分析。結(jié)果顯示,LvARMC8基因全長(zhǎng)為2917bp,其中開(kāi)放閱讀框長(zhǎng)2046 bp,編碼681個(gè)氨基酸,5′非編碼區(qū)長(zhǎng)49 bp,3′非編碼區(qū)長(zhǎng)822 bp。預(yù)測(cè)分析顯示LvARMC8編碼的蛋白質(zhì)含有6個(gè)ARM結(jié)構(gòu)域。同源性分析發(fā)現(xiàn),LvARMC8基因與內(nèi)華達(dá)古白蟻ARMC8基因的相似度最高,為71%。系統(tǒng)進(jìn)化分析結(jié)果顯示,LvARMC8和多種無(wú)脊椎動(dòng)物ARMC8聚為一支,其中與昆蟲(chóng)類(lèi)動(dòng)物赤擬谷盜、致倦庫(kù)蚊和柑橘鳳蝶的親緣關(guān)系最近。Rt-PCR分析發(fā)現(xiàn),LvARMC8基因在凡納濱對(duì)蝦多個(gè)組織中均能檢測(cè)出,在表皮中表達(dá)量最高,眼柄中表達(dá)量最低。WSSV感染后12 h,LvARMC8基因在凡納濱對(duì)蝦血液中的表達(dá)量顯著降低,但48 h后顯著升高,72 h達(dá)到最高水平。除副溶血弧菌感染后24 h外的時(shí)間點(diǎn),LvARMC8基因在凡納濱對(duì)蝦血液中的表達(dá)量均顯著升高。研究表明,LvARMC8基因可能參與凡納濱對(duì)蝦抗病免疫應(yīng)答途徑。
[Abstract]:In order to study the immune function of Armadillo-containing repeat protein 8 in the process of disease-resistant infection of Litopenaeus vannamei, the full-length c-DNA sequence of ARMC8 gene (LvARMC8,Gen Bank registration number: KX058562) of Litopenaeus vannamei was cloned by RACE-PCR technique for the first time. On-line software was used for bioinformatics analysis. The expression of LvARMC8 gene in different tissues of Penaeus vannamei and infected with white spot syndrome virus and Vibrio parahaemolyticus was analyzed by real-time fluorescence quantitative PCR (Rt-PCR). The results showed that the full length of the LvARMC8 gene was 2917bp. The open reading frame (ORF) was 2046 bp, encoding 681 amino acids, and the 5 'non coding region was 49 bp,3' noncoding region 822 bp.. Predictive analysis showed that the protein encoded by LvARMC8 contained six ARM domains. Homology analysis showed that the similarity between LvARMC8 gene and ARMC8 gene of Nevada termites was the highest (71%). Phylogenetic analysis showed that LvARMC8 and a variety of invertebrate ARMC8 were clustered into one branch, among which, the phylogenetic relationships with insect species, Cephalopheles quinquefasciatus and Citrus spp. Rt-PCR analysis showed that they were closely related to each other. LvARMC8 gene was detected in many tissues of Penaeus vannamei, the highest expression level was found in epidermis and the lowest expression was in eye stalk. After 12 h of WSSV infection, the expression of LvARMC8 gene decreased significantly in the blood of Penaeus vannamei, but increased significantly 48 h after infection, and the expression level of LvARMC8 gene in the blood of Penaeus vannamei was significantly increased after 48 h. 72 h reached the highest level. Except 24 hours after infection with Vibrio parahaemolyticus, the expression of LvARMC8 gene in the blood of Penaeus vannamei increased significantly. The results showed that LvARMC8 gene might be involved in the disease-resistant immune response pathway of Litopenaeus vannamei.
【作者單位】： 廣東海洋大學(xué)水產(chǎn)學(xué)院;南海生物資源開(kāi)發(fā)與利用協(xié)同創(chuàng)新中心;
【基金】：廣東省自然科學(xué)基金—博士啟動(dòng)項(xiàng)目(2014A030310184);廣東省自然科學(xué)基金—自由申請(qǐng)項(xiàng)目(2016A030313757) 廣東普通高校青年創(chuàng)新人才項(xiàng)目(2014KQNCX079) 廣東海洋大學(xué)優(yōu)秀青年骨干教師特別資助計(jì)劃(HDYQ2015004)~~
【分類(lèi)號(hào)】：S917.4

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