长白猪组织中牛磺酸合成关键酶及转运体的表达研究

发布时间：2018-04-15 20:08

本文选题：牛磺酸 + 半胱氨酸双加氧酶　；参考：《沈阳农业大学》2016年硕士论文

【摘要】：牛磺酸是哺乳动物体内含量最为丰富的游离氨基酸之一,生理学功能十分广泛,对生物机体器官及组织功能的发挥起着重要作用。哺乳动物体内牛磺酸的来源除自身的内源性合成外,还可通过食物摄取来获得。牛磺酸合成的关键酶有半胱氨酸双加氧酶(CDO)和半胱亚磺酸脱羧酶(CSAD/CSD), CDO-CSD途径合成牛磺酸是体内牛磺酸生物合成的重要途径,细胞需通过牛磺酸转运体(TAUT)对牛磺酸进行转运,以维持胞内牛磺酸的浓度。目前关于牛磺酸在猪体内的合成及其对猪的作用尚缺乏相关资料,本文通过RT-PCR、Western-blot和酶联免疫吸附等方法检测了牛磺酸生物合成关键酶-CDO、 CSD及转运体TAUT在猪肝脏、下丘脑、垂体、甲状腺、肾上腺、睾丸和卵巢组织中的表达以及牛磺酸的含量,探讨了这些组织中牛磺酸的来源,以期为实验室今后进一步研究牛磺酸对猪的作用及其在猪生产中的应用奠定前期基础。试验结果证明：①长白猪肝脏组织CDO cDNA编码区序列为609 bp,编码区在1-603bp,编码201个氨基酸；与GenBank中的野猪(Sus scrofa) CDO cDNA核苷酸序列同源性为99%,编码的氨基酸序列同源性为100%。②所检测的长白猪7个组织中均存在CDO、CSD、TAUTmRNA和蛋白的表达,其中CDO mRNA表达由高到低的顺序为：睾丸、卵巢、甲状腺、肝脏、垂体、下丘脑、肾上腺；CSD mRNA表达由高到低的顺序为：甲状腺、垂体、肝脏、卵巢、下丘脑、肾上腺、睾丸；TAUT mRNA表达由高到低的顺序为：垂体、肝脏、卵巢、下丘脑、甲状腺、肾上腺、睾丸。③所检测的长白猪血清和7个组织中均存在牛磺酸,且其含量由高到低为：垂体、下丘脑、肾上腺、肝脏、睾丸、卵巢、甲状腺、血清。试验结果表明：牛磺酸广泛存在于猪垂体、下丘脑、肾上腺、肝脏、睾丸、卵巢和甲状腺组织中,其来源除组织自身通过CDO-CSD途径生物合成外,还需通过TAUT从血液中摄取。牛磺酸对垂体、下丘脑、肾上腺、肝脏、睾丸、卵巢和甲状腺功能的发挥有一定的作用,但其确切作用尚需进一步研究。
[Abstract]:Taurine is one of the most abundant amino acids in mammals, the physiological function is very extensive, play an important role on biological body organs and tissue function. In addition to their own source of endogenous synthesis of taurine in mammals, can also be obtained through food intake. The key enzyme of taurine synthesis with double cysteine oxygenase the enzyme (CDO) and cysteine sulfinate decarboxylase (CSAD/CSD), CDO-CSD pathway is an important way in the synthesis of taurine taurine biosynthesis, cell by taurine transporter (TAUT) transport of taurine, to maintain the intracellular concentration of taurine. There is still a lack of relevant data in the synthesis of taurine and its effect on pig pigs in this paper, through RT-PCR, Western-blot and ELISA assay taurine biosynthesis key enzyme -CDO, CSD and TAUT in transporter Pig liver, hypothalamus, pituitary, thyroid, adrenal, testis and ovarian tissue expression and taurine content, discusses the source of taurine in these tissues, in order to further study the effect of taurine on laboratory pig and its application in pig production to lay the preliminary foundation. Test results show that the liver cells of Landrace CDO cDNA encoding sequence was 609 BP, encoding in 1-603bp, encoding 201 amino acids; and in the GenBank (Sus scrofa) CDO wild boar cDNA nucleotide sequence homology was 99%, encoding the amino acid sequences of CDO, there were the 100%. detection for the 7 Landrace tissues CSD, TAUTmRNA and protein expression the expression of CDO, mRNA in the order from high to low: testis, ovary, liver, thyroid, pituitary, hypothalamus and adrenal gland; CSD mRNA expression from high to low in the order: thyroid, pituitary, liver The hypothalamus, adrenal, ovary, testis, TAUT; expression of mRNA from high to low order: pituitary, hypothalamus, ovary, liver, thyroid, adrenal, testis. There were 3 taurine detection in serum and 7 Landrace tissues, and its content from high to low: the hypothalamus, pituitary, adrenal gland. The liver, testis, ovary, thyroid gland, serum. The results showed that taurine exists in porcine pituitary, hypothalamus, adrenal gland, liver, testis, ovary and thyroid tissue, the sources of the organization itself through the pathway of CDO-CSD biosynthesis, also need uptake from the blood by TAUT. Taurine on hypothalamus, pituitary, adrenal gland, liver, testis, ovary and thyroid function has certain effect, but its exact role needs further study.

【学位授予单位】：沈阳农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S828

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