發(fā)布時間：2018-03-12 19:36

  本文選題：分子特征　切入點：長角血蜱　出處：《畜牧獸醫(yī)學(xué)報》2017年02期 　論文類型：期刊論文

【摘要】：谷氨酰胺轉(zhuǎn)移酶(TGase)通過催化蛋白質(zhì)的谷氨酰胺殘基與賴氨酸殘基之間形成ε-(γ-谷氨�；�)賴氨酸異肽鍵,或在與肽結(jié)合的谷氨酰胺殘基處摻入伯胺,促進(jìn)蛋白質(zhì)分子內(nèi)和分子間的交聯(lián),形成網(wǎng)狀的高分子聚合物,參與多種重要的生物學(xué)活動。本研究旨在克隆和表達(dá)長角血蜱(Haemaphysalis longicornis)的谷氨酰胺轉(zhuǎn)移酶基因(HlTGase,GenBank登錄號為KX59300),并分析重組蛋白質(zhì)的活性,評估其可能的應(yīng)用價值。首先從長角血蜱上海株的成蟲提取總RNA,根據(jù)表達(dá)序列標(biāo)簽的序列信息,設(shè)計引物擴增并克隆HlTGase基因,以質(zhì)粒pPICZC為表達(dá)載體,將該基因在畢赤酵母中重組表達(dá),進(jìn)而對其編碼蛋白質(zhì)的分子特征及可能的應(yīng)用價值進(jìn)行評估。結(jié)果顯示HlTGase基因的開放閱讀框為2 262bp,編碼了一條756aa的多肽鏈,該多肽鏈具有四個谷氨酰胺轉(zhuǎn)移酶結(jié)構(gòu)域,理論相對分子質(zhì)量為84.6ku;系統(tǒng)發(fā)育樹顯示其與果蠅的谷氨酰轉(zhuǎn)移酶親緣關(guān)系最近;在畢赤酵母中成功表達(dá)的重組蛋白質(zhì)具有谷氨酰胺轉(zhuǎn)移酶的活性,能催化酪蛋白交聯(lián)成較大的分子。本研究成功地用酵母表達(dá)了長角血蜱谷氨酰胺轉(zhuǎn)移酶,重組蛋白質(zhì)有催化蛋白質(zhì)交聯(lián)的活性,具有一定的應(yīng)用前景。
[Abstract]:Transglutaminase (TGase) catalyzes the formation of 蔚-(緯 -glutamyl) lysine isopeptide bonds between glutamine residues and lysine residues of proteins, or the incorporation of primary amines at the sites of glutamine residues that bind to peptides. Promoting intramolecular and intermolecular crosslinking of proteins to form a network of polymer polymers, The aim of this study was to clone and express the glutamine transferase gene of Haemaphysalis longicornis, and to analyze the activity of recombinant protein by using GenBank accession number KX59300. To evaluate its potential application value. Firstly, total RNAs were extracted from adults of Haemaphysalis longicornis Shanghai strain. Primers were designed to amplify and clone the HlTGase gene according to the sequence information of the expressed sequence tags, and the plasmid pPICZC was used as the expression vector. The gene was expressed in Pichia pastoris, and the molecular characteristics and potential application value of the protein were evaluated. The results showed that the open reading frame of HlTGase gene was 2262 BP, encoding a 756aa polypeptide chain. The polypeptide chain has four glutamine transferase domains and the theoretical relative molecular weight is 84.6 ku.The phylogenetic tree shows that the phylogenetic tree has the closest relationship with the glutamyl transferase of Drosophila melanogaster. The recombinant protein successfully expressed in Pichia pastoris has the activity of transglutaminase and can catalyze the cross-linking of casein into larger molecules. The recombinant protein has the activity of catalyzing the cross-linking of proteins and has a certain application prospect.
【作者單位】： 福建師范大學(xué)生命科學(xué)學(xué)院福建省發(fā)育與神經(jīng)生物學(xué)重點實驗室;
【基金】：福建省自然科學(xué)基金(2014J01120) 福建師范大學(xué)生命科學(xué)學(xué)院本科生拔尖人才培養(yǎng)項目
【分類號】：S852.746

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本文編號：1603005