Human ACAT gene rs10753191 site SNP genotyping and construction of alternative splicing plasmid
Abstract: Acyl-coenzyme A: cholesterol acyltransferase (ACAT) is an intracellular enzyme catalyzing the formation of cholesteryl esters from cholesterol and long-chain fatty-acyl-coenzyme A. ACAT plays an important role in cholesterol absorption and transportation, as well as in cellular cholesterol homeostatic regulation. ACAT gene family consists of Acat1 and Acat2. In adult human tissues, ACAT1 protein is found in almost all of the cells and tissues examined, and believed to be crucial for maintaining cholesterol homeostasis. On the other hand, ACAT2 is selectively expressed in liver and small intestine, mainly involving in the dietary cholesterol absorption and assembly of apoB-containing lipoprotein. ACATs slao play important roles in some serious human diseases including atherosclerosis, Alzheimer’s disease (AD) and gallstone. For these reasons, ACAT has been considered as one of major pharmaceutical targets for developing CE-lowering, anti-atherosclerosis and/or anti-AD drugs.
Human Acyl-coenzyme A:cholesterol acyltransferase-1 (ACAT-1) cDNA K1 is distributed over two different chromosomes (chromosome 1 and 7) with two separate promoters, designated as P1 and P7.Analyzing SNPs by computer and there is an ESE at Exon10 1032.If mutatiom the C to the T , and the SR proteid will not combine on the ESE then alternative splicing happen.There is statistic shows that normal people most with the C and DHC patients with the T.My work from here.First,Klone genomic DNA inclute Exon10 from DHC patients’blood lipid.Second, sequence and analyse 1032 site.Third,use Tetra-primer ARMS-PCR to study genotyping SNPs.Nested PCR to do mutation on the basic of constructing plasmids with human ACAT1 genomic DNA.
Keywords：ACAT-1gene,DNA klone,genotyping,Nested PCR,mutation
