RESUMO
Parathyroid hormone-related protein (PTHrP) is involved in the deposition of milk calcium in mammal lactation, but its role in buffalo is unclear. In this study, the full-length coding sequence of the water buffalo PTHrP gene was first isolated using reverse transcription-polymerase chain reaction. The protein was then subjected to molecular characterization using bioinformatic methods, and the tissue expression pattern was further assayed by semi-quantitative reverse-transcription polymerase chain reaction. The water buffalo PTHrP gene contains an open reading frame of 534 base pairs encoding a polypeptide of 177 amino acid residues, a theoretical molecular weight of 20.32 kDa, and an isoelectric point of 10.00. In addition, water buffalo PTHrP was predicted to contain a signal peptide, a typical hydrophobic region with no hydrophobic transmembrane regions, and to exert its function in the cell nucleus. A conserved domain of parathyroid superfamily from amino acids 34-114 was observed in the polypeptide. Sequence comparison and the phylogenetic analysis showed that the sequence of the water buffalo PTHrP protein shared high homology with that of other mammals, particularly cattle and goat. Among the 16 tissues examined, the PTHrP gene was only expressed in adipose tissue, placenta, uterine wall, hypophysis, and mammary gland tissue, but gene expression levels were higher in the uterus wall and adipose tissue. The results of this study suggest that the PTHrP gene plays an important role in the deposition of milk calcium of water buffalo.
Assuntos
Búfalos/genética , DNA Complementar/genética , Perfilação da Expressão Gênica , Proteína Relacionada ao Hormônio Paratireóideo/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Búfalos/metabolismo , Clonagem Molecular , DNA Complementar/química , Feminino , Dados de Sequência Molecular , Proteína Relacionada ao Hormônio Paratireóideo/classificação , Proteína Relacionada ao Hormônio Paratireóideo/metabolismo , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Homologia de Sequência de AminoácidosRESUMO
OBJECTIVE: Ischemic stroke (IS) is a complex disease that resulting from the interaction of various environmental and genetic risk factors. As genetic factors exerting a direct contributory role in IS, it is one of the focus areas of identification the genetic factors of IS. This study aimed to screen bio-targets of ischemic stroke (IS), and to identify related drug molecules. MATERIALS AND METHODS: The gene expression profile GSE22255 was downloaded from Gene Expression Omnibus (GEO) database, including 20 whole blood samples from IS patients (IS group) and 20 samples from healthy controls (control group). Differentially expressed genes (DEGs) were screened out using limma package in R. Hierarchical clustering and differences between the groups analysis were conducted for confirming these DEGs. Database for Annotation, Visualization and Integrated Discovery (DAVID) and Kyoto Enrichment of Genes and Genomes (KEGG) were used to obtain the functional genes and pathways respectively. The DEGs were then entered into the WebGestalt database and related drug molecules were retrieved. RESULTS: Compared with the control group, 27 DEGs were identified from IS group including 25 up- and 2 down-regulated genes. Then functions and pathways enrichment analysis for DEGs were conducted and TNF, IL1B and TNFAIP3 were found to be both participate in apoptosis and NOD-like receptor signaling pathway. Finally, collagenase and other most-related drug molecules were identified from the DEGs. CONCLUSIONS: In addition to DEGs, several drug molecules were retrieved, which may be related with stroke. Our study provides some underlying bio-targets such as TNF, IL1B and TNFAIP3 for IS and potential drug molecules such as collagenase for the treatment of IS.