|
|
Validation of the PCR method for the determination of isotype variants in the ApoE gene
SÝKOROVÁ, Klára
Apolipoprotein E (APOE) was discovered in the early 1970s as a 34 kDa polypeptide of 299 amino acids. It plays an important role in the metabolism of cholesterol and triglycerides. It is an important component of VLDL, HDL and chylomicrons. The APOE gene is located on the long arm of chromosome 19 in region 19q13.32. It consists of three introns and four exons. It occurs in three alleles e2, e3 and e4, which encode three different isoforms. The isoforms differ in two amino acids (arginine and cysteine) at positions 112 and 158. The e3 allele is the most common, the rarer alleles e2 and e4 are associated with various types of diseases, such as cardiovascular disease, Alzheimer's disease or type III hyperlipoproteinaemia. The aim of my bachelor thesis was to write a research on the topic using information obtained from available proffesional publications, to master basic methods of molecular biology (which include DNA isolation, DNA concentration measurement, preparation of PCR reaction and gel electrophoresis) and to perform PCR method validation. The theoretical part contains general information about lipoproteins, associated with them lipoprotein metabolism and apolipoproteins. It also includes knowledge of the APOE gene, its product and the effect of APOE on various diseases. Finally, there is an overview of laboratory methods used for genetic analysis of isotype variants in the APOE gene. The practical part, which I did in the genetic laboratory GENLABS s.r.o in České Budějovice, includes the analysis of the APOE genotype and the validation of the PCR method, which was successful. Therefore, the method can be included in the daily operation of the laboratory.
|
guest ::
