Molecular Cloning,sequence Identification And Expression Profile Of Domestic Guinea Pig (Cavia Porcellus) UGT1A1, UGT1A3 And UGT1A8 Genes

Domestic guinea pig(Cavia porcellus)is an important model animal and widely used for human disease research.UDP glucuronosyltransferase 1 family,polypeptide A1(UGT1A1),UDP glucuronosyltransferase 1 family,polypeptide A3(UGT1A3)and UDP glucuronosyltransferase 1 family,polypeptide A8(UGT1A8),are three important human disease-related genes.In this project,UGT1A1,UGT1A3 UGT1A8 genes of domestic guinea pig were cloned by RT-PCR.These three genes were then deposited into GenBank database and had been assigned Accession number KT160222,KT160223 and KT160224,respectively.Subsequently,sequence analysis and tissue expression analysis were performed.The results were described as below:1.UDP glucuronosyltransferase 1 family,polypeptide A1(UGT1A1)is an important human disease-related gene.In this study,the complete coding sequence of domestic guinea pig UGT1A1 gene was amplified by RT-PCR.The open reading frame of domestic guinea pig UGT1A1 gene was 1,602 bp encoding a protein of 533 amino acids.Sequence analysis revealed that the UGT1A1 protein of domestic guinea pig shares high homology with the UGT1A1 proteins of degu(84%),damara mole-rat(84%),human(80%),northern white-cheeked gibbon(80%),colobus angolensis palliates(80%)and golden snub-nosed monkey(79%).This gene is structured in five exons and four introns as revealed by computer-assisted analysis.Results also showed that domestic guinea pig UGT1A1 genehas a close genetic relationship with the UGT1A1 gene of degu.The prediction of transmembrane helices showed that domestic guinea pig UGT1A1 might be a transmembrane protein.Expression profile analysis indicated that domestic guinea pig UGT1A1 gene was differentially expressed in detected domestic guinea pig tissues.Our experiment established primary foundation of using domestic guinea pig as model animal to study the UGT1A1-related human diseases.2.UDP glucuronosyltransferase 1 family,polypeptide A3(UGT1A3)is also a human medicine related gene.In this study,the complete coding sequence of domestic guinea pig UGT1A3 gene was amplified by RT-PCR.The open reading frame of domestic guinea pig UGT1A3 gene was 1,605 bp encoding a protein of 534 amino acids.Sequence analysis revealed that the UGT1A3 protein of domestic guinea pig shares high homology with the UGT1A3 proteins of human(75%),chimpanzee(75%),sumatran orangutan(75%),northern white-cheeked gibbon(75%),ma's night monkey(75%)and drill(74%).This gene is structured in five exons and four introns as revealed by computer-assisted analysis.Results also showed that domestic guinea pig UGT1A3 gene has a close genetic relationship with the UGT1A3 gene of ma's night monkey.The prediction of transmembrane helices revealed that domestic guinea pig UGT1A3 might be a transmembrane protein.The expression profile indicated that domestic guinea pig UGT1A3 gene was differentially expressed in detected domestic guinea pig tissues including small intestine,spleen,lung,muscle,fat,liver,heart and kidney.Our experiment established primary foundation of using domestic guinea pig as model animal to study the UGT1A3-related human diseases.3.UDP glucuronosyltransferase 1 family,polypeptide A8(UGT1A8)is another important human medicine related gene.In this study,the complete coding sequence of domestic guinea pig UGT1A8 gene was amplified by RT-PCR.The open reading frame of domestic guinea pig UGT1A8 gene was 1,596 bp encoding a protein of 531 amino acids.Sequence analysis revealed that the UGT1A8 protein of domestic guinea pig shares high homology with the UGT1A8 proteins of long-tailed chinchilla(88%),drill(84%),golden snub-nosed monkey(84%)and crab-eating macaque(83%).Results also showed thatdomestic guinea pig UGT1A8 gene has a close genetic relationship with the UGT1A8 gene of long-tailed chinchilla.The prediction of transmembrane helices revealed that domestic guinea pig UGT1A8 might be a transmembrane protein.The expression profile indicated that domestic guinea pig UGT1A8 gene was differentially expressed in detected domestic guinea pig tissues including small intestine,spleen,lung,muscle,fat,liver,heart and kidney.Our experiment established primary foundation of using domestic guinea pig as model animal to study the UGT1A8-related human diseases.