本文選題：TAUT　切入點(diǎn)：CSAD　出處：《湖南農(nóng)業(yè)大學(xué)》2016年碩士論文

【摘要】：牛磺酸對(duì)魚(yú)類生長(zhǎng)發(fā)育有調(diào)控作用,TAUT和CSAD分別是動(dòng)物細(xì)胞內(nèi)�；撬峥缒まD(zhuǎn)運(yùn)吸收的關(guān)鍵載體和自主合成的主要限速酶。本研究以鯽魚(yú)為研究對(duì)象,克隆了鯽魚(yú)TAUT和CSAD基因,檢測(cè)這兩個(gè)基因mRNA在鯽魚(yú)不同組織,晝夜節(jié)律中的相對(duì)表達(dá),同時(shí)還研究了�；撬釋�(duì)鯽魚(yú)生長(zhǎng),TAUT,CSAD表達(dá)的影響,以期為解釋�；撬釋�(duì)水產(chǎn)動(dòng)物的作用機(jī)理提供新的理論依據(jù)。1、鯽魚(yú)TAUT和CSAD基因克隆及生物信息學(xué)分析在我們的研究中,采用RACE等技術(shù),從鯽魚(yú)腸道中得到了TAUT,CSAD基因的完整的ORF序列。應(yīng)用生物信息學(xué)軟件分析序列表明：TAUT的ORF全長(zhǎng)為1978bp,編碼625個(gè)氨基酸；氨基酸區(qū)域中存在12個(gè)跨膜區(qū)域,多重序列比對(duì)分析表明鯽魚(yú)TAUT與鯉魚(yú),斑馬魚(yú)一致性高達(dá)97%。系統(tǒng)進(jìn)化分析顯示與斑馬魚(yú)的親緣關(guān)系最近,其次是鯉魚(yú)。CSAD的ORF全長(zhǎng)為1503 bp,編碼500個(gè)氨基酸；同源性分析表明,鯽魚(yú)CSAD與鯉魚(yú)CSAD的一致性高達(dá)97%；系統(tǒng)進(jìn)化分析顯示,鯽魚(yú)CSAD與鯉魚(yú)CSAD親緣關(guān)系最近。2、鯽魚(yú)TAUT和CSAD基因在不同組織和晝夜節(jié)律下的表達(dá)分析本研究采用實(shí)時(shí)熒光定量PCR技術(shù),探討TAUT和CSAD基因在鯽魚(yú)不同組織分布,晝夜節(jié)律的表達(dá)差異。結(jié)果顯示TAUT mRNA在腸組織中的相對(duì)表達(dá)量最高,顯著高于在其它組織中的表達(dá)量,在心臟和腦中的相對(duì)表達(dá)量較低。CSAD mRNA在肌肉,心臟,腸道及肝臟中的相對(duì)表達(dá)量的差異性不顯著,在腦和鰓組織中的相對(duì)表達(dá)量較低。鯽魚(yú)腸道TAUT mRNA在晝夜節(jié)律中的相對(duì)表達(dá)量呈現(xiàn)的是從0：00開(kāi)始逐漸上升到12：00,然后開(kāi)始逐漸下降的一個(gè)過(guò)程,其中12：00表達(dá)量最高,21：00的相對(duì)表達(dá)量最低。鯽魚(yú)腸道CSAD mRNA在晝夜節(jié)律中的相對(duì)表達(dá)量呈現(xiàn)的同樣是從0點(diǎn)開(kāi)始先上升后下降的過(guò)程,其中6：00時(shí)相對(duì)表達(dá)量最高,21：00的相對(duì)表達(dá)量最低。3、�；撬釋�(duì)鯽魚(yú)TAUT和CSAD基因表達(dá)的影響本試驗(yàn)以480尾(初始體質(zhì)量29.07±0.19g)鯽魚(yú)(Carassius auratus)為研究對(duì)象,隨機(jī)分成6組,每組設(shè)定4個(gè)重復(fù),每個(gè)重復(fù)有魚(yú)20尾。鯽魚(yú)基礎(chǔ)飼料中添加的�；撬岬膭┝糠謩e是0.0 g/kg、1.0g/kg、2.5 g/kg、4.0g/kg、6.0g/kg以及9.0g/kg,以0 g/kg添加組為對(duì)照組,試驗(yàn)持續(xù)8周。試驗(yàn)結(jié)果表明：(1)�；撬釋�(duì)鯽魚(yú)有促生長(zhǎng)作用,6.0 g/kg添加組的魚(yú)的增重率,特定生長(zhǎng)率,蛋白質(zhì)效率,飼料系數(shù)等都顯著高于對(duì)照組(P＜0.05)。(2)�；撬崽岣吡琐a魚(yú)血清中T3,T4的含量,添加6.0 g/kg時(shí)顯著高于對(duì)照組,且此時(shí)血清尿素氮濃度顯著低于對(duì)照組和其它處理組(P＜0.05)。(3)�；撬崮苡绊戹a魚(yú)腸道TAUT, CSAD的相對(duì)表達(dá)量。在由此可見(jiàn),適量添加�；撬崮艽龠M(jìn)鯽魚(yú)生長(zhǎng)、腸道細(xì)胞增殖及蛋白消化吸收相關(guān)基因的表達(dá)。
[Abstract]:Taurine can regulate the growth and development of fish. Taut and CSAD are the key carriers and the main rate-limiting enzymes of taurine transmembrane transport and absorption in animal cells, respectively.In this study, the TAUT and CSAD genes of crucian carp were cloned, and the relative expression of mRNA in different tissues and circadian rhythm of crucian carp was detected. The effect of taurine on the expression of TAUTCSAD in crucian carp growth was also studied.In order to provide a new theoretical basis for explaining the action mechanism of taurine on aquatic animals, the cloning of TAUT and CSAD genes and bioinformatics analysis of crucian carp were carried out using RACE and other techniques in our research.The complete ORF sequence of taut CSAD gene was obtained from the intestine of crucian carp.The results of bioinformatics software showed that the total length of ORF was 1978bp, encoding 625 amino acids, and there were 12 transmembrane regions in the amino acid region. The results of multiple sequence alignment analysis showed that the consistency of TAUT and carp was 997%.Phylogenetic analysis showed that the ORF of carp. CSAD was 1503 BP, encoding 500 amino acids. Homology analysis showed that the identity between CSAD and CSAD was 97%.Expression of TAUT and CSAD genes in different tissues and circadian Rhythm of Carassius Carassius Carassius auratus CSAD and Carp CSAD. In this study, real-time fluorescence quantitative PCR technique was used to investigate the distribution of TAUT and CSAD genes in different tissues of crucian carp.Differences in circadian rhythm expression.The results showed that the relative expression of TAUT mRNA in intestinal tissue was the highest, significantly higher than that in other tissues, and the relative expression in heart and brain was lower. CSAD mRNA was expressed in muscle and heart.There was no significant difference in the relative expression between intestine and liver, but the relative expression in brain and Gill was lower.The relative expression of TAUT mRNA in the circadian rhythm of crucian carp was gradually increased from 0:00 to 12: 00, and then decreased gradually, and the relative expression of 12:00 was the highest and the lowest at 21: 00.The relative expression of CSAD mRNA in the circadian rhythm of crucian carp intestinal tract was similar to that of rising at 0 and then decreasing.The lowest relative expression was 21: 00 at 6:00. The effect of taurine on the expression of TAUT and CSAD genes in crucian carp was studied in 480 (initial body mass 29.07 鹵0.19g) Carassius auratus.Each repeat has 20 fish.The dosage of taurine added to the basic feed of crucian carp was 0.0 g / kg ~ (-1) g / kg ~ (-1) g / kg ~ (2.5) g / kg ~ (2.5) g 路kg ~ (-1) 路kg ~ (-1) 路kg ~ (-1) 路g ~ (-1) 路kg ~ (-1), respectively.The results showed that the weight gain rate, specific growth rate, protein efficiency and feed efficiency of the group supplemented with 6. 0 g/kg of taurine were significantly higher than those of the control group (P < 0. 05, P < 0. 05, P < 0. 05, P < 0. 05). The content of T3 + T4 in the serum of Carassius auratus was significantly higher than that of the control group (P < 0. 05, P < 0. 05, P < 0. 05).The concentration of serum urea nitrogen was significantly lower than that of control group and other treatment groups (P < 0.05). Taurine could affect the relative expression of TAUTand CSAD in the intestine of crucian carp.It can be seen that appropriate amount of taurine can promote the growth of crucian carp, intestinal cell proliferation and protein digestion and absorption of related genes.
【學(xué)位授予單位】：湖南農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：S917.4;Q78

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本文編號(hào)：1714312