NCOA2 Core Promotor Methylation And Its Differential Expression Profile In Different Fat Tissue

NCOA2 belongs to Steroid receptor coactivator, which plays an important role in lipid metabolism and energy balance, and in white adipose tissue (WAT), which is used as a PPARr coactivator. Therefore, in-depth understanding of NCOA2 gene can provide a theoretical basis and technical support for the pig breeding and pig fat deposition.1. Sequence analysis of NCOA2 gene upstream regulatory region and detection of NCOA2 core promoter region.In this study, first we find its upstream regulatory region of the gene sequences from porcine published in NCOA2 NCBI database, and a variety of bio-informatics software biological analysis was carried out, and then use the dual luciferase reporter system to determine the core promoter region of the gene NCOA2. The results were as follows:NCOA2 gene 2000bp upstream of the start point of the sequence, containing CdxA, TATA, SRY, Nkx-2, GATA-X, Sox-5, AML-la, MZF1 transcriptional regulatory elements and other binding sites. Dual fluorescent reporter system testing showed, NCOA2 gene core promoter region located -483～-179.2. The methylation analysis of Pig NCOA2 gene core promoter regionDNA methylation in vertebrate is the only natural way of chemical modification on the regulation of gene expression, which plays an important role. The CpG methylation in gene promoter region, in particular in the core promoter region, can negatively regulate transcription. In this study, as the difference NCOA2 gene expression levels in subcutaneous fat tissue, perirenal fat tissue and intramuscular fat tissue. We use BSP methylation detection technology to detect NCOA2 gene core promoter region. The results were as follows:To detect the methylation level of swine NCOA2 gene core promoter region, we use BSP detection method, which found no difference among subcutaneous fat tissue, perirenal fat tissue and intramuscular fat tissue about 25 CpG sites, none has methylation, indicating NCOA2 gene differentially expression in subcutaneous fat tissue, perirenal fat tissue and intramuscular fat tissue is not relevant with its methylation level.3. SNP of NCOA2 promoter region between breedsErhualian pig was a landraces with a higher intramuscular fat content features.The experiment for the IMF differences between commercial pig(Duroc × Landrace× Yorkshire,D×L×Y) and Erhualian was analyzed to detect its relationship with NCOA2 gene expression levels, further we study the sequence of NCOA2 gene promoter region. The results were as follows:The intramuscular fat (IMF) content of Erhualian was significantly higher than that of D×L×Y crossbred pigs, the expression levels of genes of NCOA2 in Erhualian also significantly higher than commodity pigs. NCOA2 gene core promoter region -224bp the base of Erhualian is C, and commodity pig is T, which exist specific differences.