Investigation of differentially expressed transcripts in livers ofpigs

Autor: Hsiao-Ting Yang, 楊曉婷
Rok vydání: 2009
Druh dokumentu: 學位論文 ; thesis
Popis: 97
The purpose of this study was to comprehend the differentially expressed genes in the liver tissue of porcine using cDNA microarray chips. The liver tissues were collected from LD and LYD breeds. The total RNA was isolated and pooled to construct cDNA library with Creator SMART cDNA Library Construction Kit. Plasmid DNA was extracted from colonies that were random selected in cultured plates. The inserts fragments were amplified by PCR. Amplified housekeeping genes and blank solution (DMSO) were used as control for array fabrication. Total samples were collected from 8 pigs each breeds, then grouped according to sex and breed. The reverse transcription labeling and hybridization essentially followed the protocol of MICROMAXTM Direct Labeling Kit. After washing, the hybridization chips were scanned by fluorescent scanner at 532 nm for image acquisition and quantification. Data normalization, log transformation and statistics were carried out by software. The results showed that 41 genes were differentially expressed significance (P < 0.05) and observed in fold change> 1.5 with liver tissue of porcine. There are 26 differentially expressed genes and clones were found after sequencing and alignment by BLAST (GenBank), involve gamma-fibrinogen (FGG) gene, argininosuccinate synthetase 1 (ASS1) gene, beta-fibrinogen (FGB) gene, cytochrome c oxidase subunit III (COX3) gene, b-factor, properdin (BF) gene, phytanoyl-CoA 2-hydroxylase (PHYH) gene, ferritin gene, heavy polypeptide 1 (FTH1) gene, mitochondrial ribosomal protein L46 (MRPL46) gene, dehydroepiandrosterone (DHEA) gene, C-reactive protein (CRP) gene, beta-globin (HBB) gene、haptoglobin (HP) gene and ribosomal protein L19 (RPL19). These differentially expressed genes may involve in synthesis of fat and formation of obesity. Using PCR-SSCP and minisequencing to found SNPs in these sequences and analyzing the relationship between SNPs and phenotypes. ASS1 gene, BF gene and MRPL46 gene were related to pig backfat traits, and ASS1 gene, BF gene, FTH1 gene, CRP gene and PhyH gene were related to the other carcass traits. In addition. These genes may be the markers applicable to the breeding selection programs of pigs.
Databáze: Networked Digital Library of Theses & Dissertations
Externí odkaz: