Author/Editor		Jelen, Nejc
Title		Priprava in analiza DNA-mikromrež homeostaze holesterola
Type		monografija
Place		Ljubljana
Publisher		Univerza v Ljubljani, Fakulteta za kemijo in kemijsko tehnologijo
Publication year		2004
Volume		str. 43
Language		slo
Abstract		Cardiovascular disease is amongst the most common causes of death in the developed world. Factors contributing to these conditions are both genetic and environmental. However, their interaction is difficult to describe using classical molecular biology approaches. The modern approach applies functional genomics with gene expression studies (analysis of the transcriptome using DNA microarrays), protein and metabolite studies, enabling a holistic insight into physiological as well as pathophysiological states of the cell and the organism. DNA microarrays represent a relatively new technology, which enables simultaneous analysis of expression of up to tens of thousands of genes. This opens a new, holistic line of approach for study of function, regulation and interaction of a large number of genes, far surpassing the usual hybridisation techniques. Several kinds of DNA microarrays east, differing primarily in the type of the DNA that is immobilised on the microarray (either cDNA or oligonucleotides) and in the method of application the DNA onto the array, which can be done robotically by using pre-synthesised DNA or in situ synthesis of DNA directly on the microarray). Microarrays using cDNA, where a part or the whole cDNA of the studied genes is immobilised, are particularly suited for use in academic institutions (i.e. in-house microarrays) due to their flexibility and relatively low costs. For the purpose of this thesis, specific probes were produced for the STEROTALK cDNA microarray containing immobilised fragments of cDNA genes, which are responsible for cholesterol homeostasis. Using polymerase chain reaction with probe specific primer oligonucleotides, we amplified the gene fragments from tissue-specific cDNA libraries, cloned them and checked the nucleotide sequence. (Abstract truncated at 2000 characters)
Descriptors		CARDIOVASCULAR DISEASES CHOLESTEROL OLIGONUCLEOTIDE PROBES POLYMERASE CHAIN REACTION