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

【摘要】：�；撬釋︳~類生長發(fā)育有調控作用,TAUT和CSAD分別是動物細胞內�；撬峥缒まD運吸收的關鍵載體和自主合成的主要限速酶。本研究以鯽魚為研究對象,克隆了鯽魚TAUT和CSAD基因,檢測這兩個基因mRNA在鯽魚不同組織,晝夜節(jié)律中的相對表達,同時還研究了�；撬釋a魚生長,TAUT,CSAD表達的影響,以期為解釋�；撬釋λa動物的作用機理提供新的理論依據。1、鯽魚TAUT和CSAD基因克隆及生物信息學分析在我們的研究中,采用RACE等技術,從鯽魚腸道中得到了TAUT,CSAD基因的完整的ORF序列。應用生物信息學軟件分析序列表明：TAUT的ORF全長為1978bp,編碼625個氨基酸；氨基酸區(qū)域中存在12個跨膜區(qū)域,多重序列比對分析表明鯽魚TAUT與鯉魚,斑馬魚一致性高達97%。系統(tǒng)進化分析顯示與斑馬魚的親緣關系最近,其次是鯉魚。CSAD的ORF全長為1503 bp,編碼500個氨基酸；同源性分析表明,鯽魚CSAD與鯉魚CSAD的一致性高達97%；系統(tǒng)進化分析顯示,鯽魚CSAD與鯉魚CSAD親緣關系最近。2、鯽魚TAUT和CSAD基因在不同組織和晝夜節(jié)律下的表達分析本研究采用實時熒光定量PCR技術,探討TAUT和CSAD基因在鯽魚不同組織分布,晝夜節(jié)律的表達差異。結果顯示TAUT mRNA在腸組織中的相對表達量最高,顯著高于在其它組織中的表達量,在心臟和腦中的相對表達量較低。CSAD mRNA在肌肉,心臟,腸道及肝臟中的相對表達量的差異性不顯著,在腦和鰓組織中的相對表達量較低。鯽魚腸道TAUT mRNA在晝夜節(jié)律中的相對表達量呈現的是從0：00開始逐漸上升到12：00,然后開始逐漸下降的一個過程,其中12：00表達量最高,21：00的相對表達量最低。鯽魚腸道CSAD mRNA在晝夜節(jié)律中的相對表達量呈現的同樣是從0點開始先上升后下降的過程,其中6：00時相對表達量最高,21：00的相對表達量最低。3、�；撬釋a魚TAUT和CSAD基因表達的影響本試驗以480尾(初始體質量29.07±0.19g)鯽魚(Carassius auratus)為研究對象,隨機分成6組,每組設定4個重復,每個重復有魚20尾。鯽魚基礎飼料中添加的�；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添加組為對照組,試驗持續(xù)8周。試驗結果表明：(1)�；撬釋a魚有促生長作用,6.0 g/kg添加組的魚的增重率,特定生長率,蛋白質效率,飼料系數等都顯著高于對照組(P＜0.05)。(2)�；撬崽岣吡琐a魚血清中T3,T4的含量,添加6.0 g/kg時顯著高于對照組,且此時血清尿素氮濃度顯著低于對照組和其它處理組(P＜0.05)。(3)牛磺酸能影響鯽魚腸道TAUT, CSAD的相對表達量。在由此可見,適量添加牛磺酸能促進鯽魚生長、腸道細胞增殖及蛋白消化吸收相關基因的表達。
[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.
【學位授予單位】：湖南農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：S917.4;Q78

【參考文獻】

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1 徐亞沙;李書俊;吳芹;陸遠富;劉杰;;小鼠肝臟膽酸合成及轉運基因表達的晝夜節(jié)律[J];遵義醫(yī)學院學報;2015年05期

2 張小龍;鄧歡;陳澄;張素云;唐志如;;�；撬徂D運體的調節(jié)機制及其在動物營養(yǎng)研究中的意義[J];動物營養(yǎng)學報;2013年10期

3 周玲;王曉清;劉臻;馮軍廠;郝光;孫浪;于U，

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