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Cloning,Chromosome Mapping and Functional Study of Pig SREBPs and CREB-1

Author: NieTao
Tutor: YangZaiQing
School: Huazhong Agricultural University
Course: Biochemistry and Molecular Biology
Keywords: SREBPs CREB - 1 Cloning Chromosomal localization Tissue distribution Expression and regulation
CLC: Q953
Type: Master's thesis
Year: 2005
Downloads: 179
Quote: 1
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Abstract


The sterol regulatory element binding protein (sterol regulatory element binding Proteins, SREBPs) SREBPs and cAMP response element binding protein (cAMP response element binding protein) CREB-1 is an important transcription factor in vivo, SREBPs major regulation of fatty acid and cholesterol synthesis, CREB-1 has a broader role. The SREBPs family three SREBP proteins: SREBP-1a, SREBP-1c and SREBP-2. People, SREBP-2 gene on chromosome 22 encoding. SREBP-1a and SREBP-1c derived from the same gene on chromosome 17, and select the type of the promoter of this gene to produce two different first exon, but the remaining exons. SREBP-1, 2 function have focused on SREBP-1c tend to activate the synthesis of fatty acids, and SREBP-2 tend to activate the synthesis of cholesterol. SREBP-1a is constitutively expressed, but capable of acting on all genes containing SRE sites. This study has successfully obtained swine part of the coding sequence of SREBP-2 and CREB-1, the entire coding sequence. Pig SREBP-2 with human Chinese hamster SREBP-2 nucleotide sequence homology is 86.4% and 90.9%, respectively: the homology of the nucleotide sequence of CREB-1, CREB-1 and norvegicus 95.1% and 96.8%, respectively. Announced SREBP-1c sequence analysis of SREBP-1c and SREBP-1c and Mus musculus nucleotide sequence homology is 88.5% and 78.8%, respectively. Be seen in the evolution of these three genes are very conservative. Radiation hybrid panel technology, SREBP-2 is positioned at the ends of chromosomes, 5 of the pig. The experimental results are compared with the man-pig gene mapping results are consistent. By semi-quantitative PCR method, beta-actin gene as an internal study, on SREBPs and CREB-1 distribution in the Meishan ten different organizations. Experimental results show that the three genes are widely expressed, SREBP-1, the highest level of expression in adipose tissue, SREBP-2 in the highest amount of liver tissue expression and CREB-1 tend expressed in the brain tissue. - Fat tissue, the expression of SREBP-1 is higher than that of SREBP-2 in the pig lipids generated organ. In addition, the use of the gene transfection and drug stimulation method for the the SREBPs regulating the expression of the study. COS7 cells with beta-stimulants stimulate CREB-1 expression level rise, SREBPs expression levels decline, especially the major lipid synthesis decreased expression of SREBP-1 is very obvious. The experimental results show that beta-doping through SREBPs factor, especially by the SREBP-1 to achieve the role of inhibition of fatty acid synthesis. CREB-2 is activated

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