豚鼠UGT1A1、UGT1A3和UGT1A8基因的克

发布时间：2018-06-04 15:34

本文选题：豚鼠 + UGT1A1　；参考：《云南农业大学》2017年硕士论文

【摘要】：实验动物与人类医学发展息息相关,豚鼠作为实验动物,在人类医学研究中起重要作用,UGT1A1、UGT1A3和UGT1A8是人类疾病相关的重要基因。本研究以豚鼠实验材料,通过RT-PCR方法克隆了豚鼠的UGT1A1基因(GenBank登录号:KT160222)、UGT1A3基因(GenBank登录号:KT160224)和UGT1A8基因(GenBank登录号:KT160223),并在此基础上,依据生物信息学工具分析了三个基因的核苷酸及氨基酸序列特性。利用实时荧光定量PCR分析,对这三个基因在豚鼠不同组织部位的表达情况进行了分析。主要实验结果如下:1.UDP-葡萄糖醛酸转移酶1家族多肽A1(UDP glucuronosyltransferase 1family,polypeptide A1,UGT1A1)是重要的人类疾病相关基因,本研究采用RT-PCR的方法获得UGT1A1基因cDNA序列全长,该基因开放阅读框全长为1602bp,编码533个氨基酸,该基因编码蛋白与Degu、达马拉鼹鼠、人、北部白颊长臂猿、疣猴、川金丝猴的UGT1A1蛋白分别具有84%、84%、80%、80%、80%、79%的高度同源性。计算机辅助分析表明该基因由五个外显子和四个内含子构成。进化分析表明,豚鼠UGT1A1基因与Degu UGT1A1基因具有较近的亲缘关系。跨膜螺旋预测显示,豚鼠UGT1A1可能是一种跨膜蛋白。组织表达谱分析表明,豚鼠UGT1A1基因在豚鼠各组织中差异表达。本研究为采用豚鼠作为模式动物研究人类UGT1A1基因相关疾病奠定了分子基础。2.UDP-葡萄糖醛酸转移酶1家族多肽A3(UDP glucuronosyltransferase 1family,polypeptide A3,UGT1A3)是另外一个重要的人类医学的相关基因。本研究利用RT-PCR获得UGT1A3基因cDNA序列全长,该基因开放阅读框全长为1605bp,编码534个氨基酸,该基因编码蛋白与人类、黑猩猩、苏门答腊猩猩、北方白颊长臂猿、马夜猴和鬼狒的UGT1A3蛋白分别具有75%、75%、75%、75%、75%、74%的高度同源性。计算机辅助分析表明该基因由五个外显子和四个内含子构成。进化分析表明,豚鼠UGT1A3基因与马夜猴UGT1A3基因具有较近的亲缘关系。跨膜螺旋预测显示,豚鼠UGT1A3是一种跨膜蛋白。组织表达谱分析表明豚鼠UGT1A3基因在小肠、脾、肺、肌肉、脂肪、肝脏、心脏和肾脏等组织中差异表达。本研究为采用豚鼠作为模式动物研究UGT1A3基因相关的人类疾病奠定了分子基础。3.UDP-葡萄糖醛酸转移酶1家族多肽A8(UDP glucuronosyltransferase 1family,polypeptide A8,UGT1A8)也是重要人类疾病相关基因。本研究采用RT-PCR获得UGT1A8基因cDNA全长序列,该基因开放阅读框全长为1596bp,编码531个氨基酸,该基因编码蛋白与长尾龙猫、鬼狒、川金丝猴、食蟹猕猴的UGT1A8蛋白分别具有88%、84%、84%、83%的高度同源性。进化分析表明,豚鼠UGT1A8基因与长尾龙猫UGT1A8基因具有较近的亲缘关系。跨膜螺旋预测显示,豚鼠UGT1A8是一种跨膜蛋白。组织表达谱分析表明豚鼠UGT1A8基因在小肠、脾、肺、肌肉、脂肪、肝脏、心脏和肾脏等组织中差异表达。本研究为采用豚鼠作为模式动物研究UGT1A8基因相关人类疾病奠定了分子基础。
[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.
【学位授予单位】：云南农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S865.11

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