丁詩同臺灣大學：畜產學研究所柯雅慧Ko, Ya-HueiYa-HueiKo2007-11-292018-06-292007-11-292018-06-292006http://ntur.lib.ntu.edu.tw//handle/246246/63622本試驗目的為探討固醇調節區域結合蛋白（Sterol regulatory element binding protein, SREBP）在雞隻中對於脂質新生成及膽固醇合成代謝基因的調控功能。本試驗分為兩部分進行，試驗一：將16隻7週齡肉雞隨機分成兩組，每組8隻，一組禁食24小時（禁食組），一組禁食24小時後給予任食5小時（飽食組），以即時定量聚合To study the role of sterol regulatory element binding-proteins (SREBPs) in lipogenesis and cholesterol synthesis in the chicken, two experiments were carried out. First, 16 seven-week-old broilers were allocated into 2 groups treated with either fasted for 24 h or refed for 5 h after a 24 h-fast. The mRNA concentrations for SREBPs and other lipogenic genes in the liver were determined by quantitative real-time PCR. The hepatic mRNA relative abundance of lipogenic genes and genes in cholesterol synthesis were significantly greater (p<0.001) in the refed than the fasted broilers. Similar results were demonstrated with northern analysis technique. The data suggest that in the livers of fasted broilers, the lipogenesis and cholesterogenesis activities were inhibited. The data also demonstrated that the expression of lipogenic genes coordinate well as a group during the refed period. Second, three small interfering RNA(siRNA) oligonucleotides against the SREBP1 were designed to be used in transfecting chicken hepatocarcinoma cell line LMH. One of the 3 siRNAs effectively reduced SREBP1 mRNA concentration (p<0.01). The acetyl coenzyme A carboxylase目次 中文摘要 1 英文摘要 2 緒言 3 一、固醇調節區域結合蛋白 6 二、核糖核酸干擾 10 試驗目的 13 試驗一、雞隻禁食-飽食試驗 14 結果 21 討論 27 試驗二、利用肝細胞培養測定SREBP之功能 29 結果 34 討論 37 總結 39 參考文獻 40 附錄一、組織及細胞RNA之萃取 45 附錄二、甲醛變性膠體電泳 48 附錄三、反轉錄反應 48 附錄四、LMH細胞培養 50 附錄五、siRNA轉染 52 表次 表一、即時聚合3046360 bytesapplication/pdfen-US雞隻肝臟固醇調結區域結合蛋白脂質新生成chickenliversterol regulatory element binding proteinlipogenesisthesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/63622/1/ntu-95-R92626012-1.pdf