本文選題：斜帶石斑魚 + �；撬徂D(zhuǎn)運(yùn)載體��； 參考：《集美大學(xué)》2016年碩士論文

【摘要】：石斑魚(Epinephelus)是一種經(jīng)濟(jì)價(jià)值很高的暖水性海水魚類,在我國福建、廣東、海南等沿海地區(qū)養(yǎng)殖規(guī)模很大。石斑魚�；撬岷铣赡芰^弱,主要靠外源食物供給來滿足其生長發(fā)育的需要。�；撬徂D(zhuǎn)運(yùn)蛋白(taurine transporter,TauT)是牛磺酸跨膜轉(zhuǎn)運(yùn)的重要載體,在魚類的生長、代謝及性成熟過程中發(fā)揮著重要作用。本文克隆了斜帶石斑魚TauT基因及飼料中不同牛磺酸水平對斜帶石斑魚(Epinephelus coioides)組織TauT mRNA表達(dá)的影響。主要研究結(jié)果如下:1、斜帶石斑魚TauT基因克隆首次從斜帶石斑魚肝臟中分離得到TauT基因編碼區(qū)序列全長,長度為1881bp,編碼626個(gè)氨基酸,其分子質(zhì)量為70126.80Da,等電點(diǎn)為7.834。通過對其氨基酸序列分析發(fā)現(xiàn),斜帶石斑魚TauT具有12個(gè)跨膜區(qū)域,且在第三跨膜區(qū)域與第四跨膜區(qū)域之間存在這一個(gè)親水基環(huán),這個(gè)環(huán)含有三個(gè)潛在的N-糖基化位點(diǎn)。通過同源性分析和系統(tǒng)發(fā)育分析,斜帶石斑魚TauT基因與鱸魚親緣關(guān)系最近,其次為大西洋鮭,再次為羅非魚,與其斑馬魚和鯉魚等其它淡水魚類親緣關(guān)系較遠(yuǎn),而與鳥、哺乳類的親緣關(guān)系則更遠(yuǎn)。這符合傳統(tǒng)分類學(xué)意義。2、組織表達(dá)分析檢測了斜帶石斑魚不同組織的TauT mRNA表達(dá)水平。結(jié)果發(fā)現(xiàn),TauT mRNA在所有被測組織中都有表達(dá),但在不同組織的表達(dá)水平不一致,以肝臟、心臟、腦表達(dá)水平最高,腎臟、鰓、脾臟組織的表達(dá)次之,以腸道、肌肉、脂肪組織的表達(dá)水平為最低。3、飼料�；撬釋π睅唪~生長的影響本實(shí)驗(yàn)配制牛磺酸含量為0(D1)、0.5%(D2)、1%(D3)、1.5%(D4)的4種試驗(yàn)飼料,飼喂斜帶石斑魚84d,并分別于飼喂的第28d、56d、84d稱重和采樣。結(jié)果表明,無論飼喂的第28d、第56d還是第84d,牛磺酸添加組魚的增重率均顯著高于對照組,且1%�；撬崽砑咏M增重率顯著高于其他(p0.05)。隨著飼料�；撬崴降纳�,斜帶石斑魚肝臟、肌肉、腸道牛磺酸含量顯著增高(p0.05),且隨著養(yǎng)殖d數(shù)的增加而增加(p0.05)。4、飼料�；撬釋π睅唪~TauT mRNA表達(dá)的影響在飼喂的第28d時(shí),牛磺酸添加組魚各個(gè)組織的TauT mRNA相對表達(dá)量顯著高于對照組。D3、D4組的TauT mRNA相對表達(dá)量顯著高于D1、D2組(p0.05),但D3組的表達(dá)量與D4組差異不顯著(p0.05)。飼喂的第56d、84d各組TauT相對表達(dá)量呈現(xiàn)出與飼喂第28d時(shí)一致的結(jié)果。同一組織TauT mRNA的相對表達(dá)量隨著飼喂時(shí)間的增加呈現(xiàn)逐漸下降的趨勢。
[Abstract]:Epinephelus (Epinephelus) is a kind of warm-water marine fish with high economic value. It is cultivated on a large scale in the coastal areas of Fujian, Guangdong and Hainan in China. The ability of taurine synthesis of grouper is weak, which mainly depends on exogenous food supply to meet the needs of its growth and development. Taurine transporter TauTis is an important carrier of taurine transmembrane transport, which plays an important role in the growth, metabolism and sexual maturation of fish. In this paper, we cloned the TauT gene of Epinephelus coioides and the effect of different taurine levels on the expression of TauT mRNA in the tissues of Epinephelus coioides. The main results are as follows: 1. The TauT gene coding region was cloned from the liver of the oblique grouper for the first time. The length of the coding region was 1881 BP, encoding 626 amino acids, with molecular weight of 70126.80 Daand isoelectric point of 7.834. The amino acid sequence analysis showed that TauT had 12 transmembrane regions, and there was a hydrophilic ring between the third and fourth transmembrane regions, which contained three potential N-glycosylation sites. By homology analysis and phylogenetic analysis, TauT gene was closely related to perch, followed by Atlantic salmon, tilapia and other freshwater fishes such as zebrafish and carp. Mammals are more closely related. This was in accordance with the traditional taxonomic significance. 2. The expression level of TauT mRNA in different tissues of grouper was detected by tissue expression analysis. The results showed that TauT mRNA was expressed in all the tested tissues, but the expression level was different in different tissues. The highest expression level was found in liver, heart and brain, followed by the expression in kidney, Gill, spleen, intestinal tract and muscle. The effect of feed taurine on the growth of the oblique grouper was the lowest in adipose tissue. The experiment was conducted to prepare four experimental diets with taurine content of 0 D _ (1) and 0.5 ~ (0.5) D _ (2) and D _ (3) and 1.5 ~ (5) D _ (4). The four experimental diets were fed to the oblique grouper for 84 days and were weighed and sampled on the 28th day, 56d ~ (84) d, respectively. The results showed that the weight gain rate of taurine added group was significantly higher than that of control group on the 28th day, 56th day and 84th day, and the weight gain rate of 1% taurine group was significantly higher than that of other groups. The content of taurine in liver, muscle and intestine increased significantly with the increase of feed taurine level, and increased with the increase of the number of cultured days. The effect of feed taurine on the expression of TauT mRNA in oblique grouper was 28 days after feeding. The relative expression of TauT mRNA in all tissues of taurine group was significantly higher than that in control group. D3 group was significantly higher than that in D1 D 2 group (P 0.05), but the expression of TauT mRNA in D3 group was not significantly different from that in D 4 group (P 0.05). The relative expression of TauT in each group on the 56th day after feeding showed the same result as that on the 28th day. The relative expression of TauT mRNA in the same tissue decreased with the increase of feeding time.
【學(xué)位授予單位】：集美大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S917.4

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