本文選題：大黃魚 + 鰻弧菌　； 參考：《水產(chǎn)學(xué)報》2017年05期

【摘要】：為探究過氧化氫酶(catalase,CAT)對大黃魚機體的保護作用,實驗基于大黃魚基因組序列數(shù)據(jù)庫克隆獲得CAT基因1584 bp的完整開放閱讀框(GenBank登陸號:KKF-14425.1),該序列編碼527個氨基酸殘基,包含與其他動物高度保守的酶活性中心序列FDRERIPERVVHAKGA、亞鐵血紅素結(jié)合信號序列RLFSYPDTH、3個催化位點殘基His75、Asn148和Tyr358,以及12個NADPH結(jié)合位點等,理論分子量為59.98 ku,等電點為8.37。多序列比對顯示,大黃魚CAT氨基酸序列與其他魚類具有較高的一致性,與同屬于石首魚科的軍曹魚和條石鯛同源性高達94%,在進化樹中也聚類于同一進化分支,說明該序列為CAT家族成員。實時熒光定量PCR檢測顯示,大黃魚CAT基因在所檢測的7種組織(肝臟、脾臟、腦、腎、肌肉、鰓、腸)中均有表達,但在肝臟中表達水平最高(為肌肉中的6.68倍)。鰻弧菌感染后,大黃魚肝組織中CAT基因的表達隨著時間的推移而變化明顯,感染后12 h,達到最高(7.48倍),隨后逐漸下降,到72 h已基本恢復(fù)到原始水平,注射PBS的對照組,CAT基因表達只略有上調(diào),說明病原菌侵染可能引起魚體產(chǎn)生大量活性氧自由基及H2O2,CAT基因則可以清除體內(nèi)過量的活性氧,進而防止它們對細胞造成損傷。
[Abstract]:In order to investigate the protective effect of catalase (catalase) on large yellow croaker, the complete open reading frame of CAT gene (1584 BP) was obtained based on the genomic sequence database of Pseudosciaena Crocea, which encodes 527 amino acid residues. It contains highly conserved enzyme activity center sequence FDRERIPERVHAKGA, heme hemin binding signal sequence RLFSYPDTH3, three catalytic sites His75 An 148 and Tyr358, and 12 NADPH binding sites. The theoretical molecular weight is 59.98 kuand the isoelectric point is 8.37. Multiple sequence alignment showed that the amino acid sequence of CAT of Pseudosciaena Crocea was highly consistent with other fishes, and had a high homology with the codon and Sparus species belonging to the same family, and was also clustered in the same evolutionary branch in the evolutionary tree, and the amino acid sequence of Pseudosciaena Crocea was similar to that of other fishes. The sequence is a member of CAT family. Real-time fluorescence quantitative PCR analysis showed that CAT gene was expressed in 7 tissues (liver, spleen, brain, kidney, muscle, Gill and intestine), but the highest expression level was found in liver (6.68 times as high as that in muscle). After infection with Vibrio anguilli, the expression of CAT gene in liver tissue of large yellow croaker changed obviously with the passage of time. At 12 hours after infection, the expression of CAT gene reached the highest level of 7.48 times, then decreased gradually, and returned to its original level at 72 h. The expression of cat gene in the control group injected with PBS was only slightly up-regulated, which indicated that the pathogen infection might cause a large number of reactive oxygen free radicals and H _ 2O _ 2 cat gene could scavenge excessive reactive oxygen species in the body and prevent them from damaging the cells.
【作者單位】： 浙江海洋大學(xué)國家海洋設(shè)施養(yǎng)殖工程技術(shù)研究中心;
【基金】：國家自然科學(xué)基金(41606148) 浙江省自然科學(xué)基金(LY14C190004) 浙江省大學(xué)生科技創(chuàng)新活動計劃暨新苗人才計劃(2015R411005) 舟山市科技局項目(2015C41014)~~
【分類號】：S943

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