本文選題：豚鼠 + UGT1A1��； 參考：《云南農(nóng)業(yè)大學》2017年碩士論文

【摘要】：實驗動物與人類醫(yī)學發(fā)展息息相關(guān),豚鼠作為實驗動物,在人類醫(yī)學研究中起重要作用,UGT1A1、UGT1A3和UGT1A8是人類疾病相關(guān)的重要基因。本研究以豚鼠實驗材料,通過RT-PCR方法克隆了豚鼠的UGT1A1基因(GenBank登錄號:KT160222)、UGT1A3基因(GenBank登錄號:KT160224)和UGT1A8基因(GenBank登錄號:KT160223),并在此基礎(chǔ)上,依據(jù)生物信息學工具分析了三個基因的核苷酸及氨基酸序列特性。利用實時熒光定量PCR分析,對這三個基因在豚鼠不同組織部位的表達情況進行了分析。主要實驗結(jié)果如下:1.UDP-葡萄糖醛酸轉(zhuǎn)移酶1家族多肽A1(UDP glucuronosyltransferase 1family,polypeptide A1,UGT1A1)是重要的人類疾病相關(guān)基因,本研究采用RT-PCR的方法獲得UGT1A1基因cDNA序列全長,該基因開放閱讀框全長為1602bp,編碼533個氨基酸,該基因編碼蛋白與Degu、達馬拉鼴鼠、人、北部白頰長臂猿、疣猴、川金絲猴的UGT1A1蛋白分別具有84%、84%、80%、80%、80%、79%的高度同源性。計算機輔助分析表明該基因由五個外顯子和四個內(nèi)含子構(gòu)成。進化分析表明,豚鼠UGT1A1基因與Degu UGT1A1基因具有較近的親緣關(guān)系�？缒ぢ菪A(yù)測顯示,豚鼠UGT1A1可能是一種跨膜蛋白。組織表達譜分析表明,豚鼠UGT1A1基因在豚鼠各組織中差異表達。本研究為采用豚鼠作為模式動物研究人類UGT1A1基因相關(guān)疾病奠定了分子基礎(chǔ)。2.UDP-葡萄糖醛酸轉(zhuǎn)移酶1家族多肽A3(UDP glucuronosyltransferase 1family,polypeptide A3,UGT1A3)是另外一個重要的人類醫(yī)學的相關(guān)基因。本研究利用RT-PCR獲得UGT1A3基因cDNA序列全長,該基因開放閱讀框全長為1605bp,編碼534個氨基酸,該基因編碼蛋白與人類、黑猩猩、蘇門答臘猩猩、北方白頰長臂猿、馬夜猴和鬼狒的UGT1A3蛋白分別具有75%、75%、75%、75%、75%、74%的高度同源性。計算機輔助分析表明該基因由五個外顯子和四個內(nèi)含子構(gòu)成。進化分析表明,豚鼠UGT1A3基因與馬夜猴UGT1A3基因具有較近的親緣關(guān)系�？缒ぢ菪A(yù)測顯示,豚鼠UGT1A3是一種跨膜蛋白。組織表達譜分析表明豚鼠UGT1A3基因在小腸、脾、肺、肌肉、脂肪、肝臟、心臟和腎臟等組織中差異表達。本研究為采用豚鼠作為模式動物研究UGT1A3基因相關(guān)的人類疾病奠定了分子基礎(chǔ)。3.UDP-葡萄糖醛酸轉(zhuǎn)移酶1家族多肽A8(UDP glucuronosyltransferase 1family,polypeptide A8,UGT1A8)也是重要人類疾病相關(guān)基因。本研究采用RT-PCR獲得UGT1A8基因cDNA全長序列,該基因開放閱讀框全長為1596bp,編碼531個氨基酸,該基因編碼蛋白與長尾龍貓、鬼狒、川金絲猴、食蟹獼猴的UGT1A8蛋白分別具有88%、84%、84%、83%的高度同源性。進化分析表明,豚鼠UGT1A8基因與長尾龍貓UGT1A8基因具有較近的親緣關(guān)系�？缒ぢ菪A(yù)測顯示,豚鼠UGT1A8是一種跨膜蛋白。組織表達譜分析表明豚鼠UGT1A8基因在小腸、脾、肺、肌肉、脂肪、肝臟、心臟和腎臟等組織中差異表達。本研究為采用豚鼠作為模式動物研究UGT1A8基因相關(guān)人類疾病奠定了分子基礎(chǔ)。
[Abstract]:Experimental animals are closely related to the development of human medicine. As experimental animals, guinea pigs play an important role in human medical research. UGT1A1A3 and UGT1A8 are important genes related to human diseases. In this study, the UGT1A1 gene of guinea pig was cloned from guinea pigs by RT-PCR method. The accession number of GenBank was: KT160222UGT1A3 (GenBank accession number: KT160224) and UGT1A8 gene (GenBank accession number: KT160223). The nucleotide and amino acid sequences of the three genes were analyzed by bioinformatics tools. The expression of these three genes in different tissues of guinea pigs was analyzed by real-time fluorescence quantitative PCR analysis. The main results are as follows: 1.UDP- glucuronyltransferase 1 family polypeptide A1(UDP glucuronosyltransferase 1 family polypeptide A1UGT1A1) is an important human disease-related gene. In this study, the cDNA sequence of UGT1A1 gene was obtained by RT-PCR. The open reading frame of this gene is 1602bpand encodes 533 amino acids. The UGT1A1 protein of this gene is highly homologous with that of Deguo, Damara Mole, human, northern cheekbone gibbon, verrucous monkey and Sichuan golden monkey, respectively. Computer aided analysis showed that the gene consisted of five exons and four introns. Evolutionary analysis showed that there was a close relationship between UGT1A1 gene and Degu UGT1A1 gene in guinea pigs. Transmembrane helix prediction showed that guinea pig UGT1A1 may be a transmembrane protein. Tissue expression profile analysis showed that guinea pig UGT1A1 gene was differentially expressed in different tissues of guinea pig. This study provides a molecular basis for the study of human UGT1A1 gene-related diseases using guinea pigs as a model animal. 2.UDP- glucuronyltransferase 1 family polypeptide A3UGT1A3) is another important gene related to human medicine. In this study, the cDNA sequence of UGT1A3 gene was obtained by RT-PCR. The open reading frame of this gene was 1605 BP, encoding 534 amino acids. The gene encodes a protein similar to that of human, chimpanzee, Sumatran gorilla and gibbon. The UGT1A3 protein of the horseshoe monkey and the monkey has a high homology of 75% and 74% respectively. Computer aided analysis showed that the gene consisted of five exons and four introns. The phylogenetic analysis showed that there was a close relationship between guinea pig UGT1A3 gene and horse night monkey UGT1A3 gene. Transmembrane helix prediction showed that guinea pig UGT1A3 was a transmembrane protein. Tissue expression profile analysis showed that UGT1A3 gene was differentially expressed in small intestine, spleen, lung, muscle, fat, liver, heart and kidney. This study provides a molecular basis for the study of human diseases associated with UGT1A3 gene using guinea pigs as model animals. 3. UDP- glucuronyltransferase 1 family polypeptide A8(UDP glucuronosyltransferase 1 family polypeptide A8 (UGT1A8) is also an important human disease-related gene. In this study, the full-length cDNA sequence of UGT1A8 gene was obtained by RT-PCR. The open reading frame of this gene was 1596bp, encoding 531 amino acids. The UGT1A8 protein of Rhesus macaques has a high homology of 88.3%. Evolutionary analysis showed that the relationship between guinea pig UGT1A8 gene and UGT1A8 gene was close. Transmembrane helix prediction showed that guinea pig UGT1A8 was a transmembrane protein. Tissue expression profile analysis showed that UGT1A8 gene was differentially expressed in small intestine, spleen, lung, muscle, fat, liver, heart and kidney. This study provides a molecular basis for the study of human diseases associated with UGT1A8 gene using guinea pigs as model animals.
【學位授予單位】：云南農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S865.11

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