Molecular biology in arteriosclerosis research.

Abstract

The topics discussed In this article illustrate how molecular biology will have a dramatic Impact on arteriosclerosis research. DNA clones for a small number of relevant proteins have been isolated, and studies are underway in numerous laboratories to extend these initial studies. The techniques of molecular biology will provide major advances in our understanding of numerous proteins directly or indirectly involved in the atherogenlc process. Cloning technology will solve the primary structures of many proteins that can not be purified In quantities sufficient for classical methods of analysis. Studies of regulation will benefit from the availability of DNA probes, the ability to generate site-directed antibodies, and the use of reverse genetics to Identify nucleic acid sequences involved in the regulation of gene expression. Studies of gene structure and genetic polymorphisms will unravel the genetic basis for defects In llpld and lipoprotein metabolism and should provide valuable reagents for clinical screening and diagnosis. The reverse genetics approach will permit the systematic analysis of structure-function relationships at the protein level In a manner not previously possible. Each of these will contribute to our understanding of the atherogenic process and should provide insight into ways of preventing and treating arteriosclerosis. T he purpose of this article is to explore how molecular biology can have an impact on research in the broad field of arteriosclerosis. The past few years have seen the techniques of molecular biology applied to an increasingly wider range of problems directly or indirectly related to arteriosclerosis. The first focus of research in this area has been the isolation and characterization of complementary DNA (cDNA) clones for the major apolipoproteins of the plasma lipoproteins. The apolipoproteins were prime targets for initial studies because they are abundant products of lipoprotein-producing tissues and the amino acid sequences of many of these proteins are known. While the current literature seems filled with cloning papers, we only have scratched the surface

Gene expression in HCV-associated HCC-identification of an upregulated gene encoding a protein related to ubiquitin conjugating enzyme

“…In extreme cases all the RNA will migrate as diffuse smear of low molecular weight. This is frequently caused by inadequate de-ionisation of the glyoxal (Williams, 1985). Alternatively, it may arise from RNase contamination in one of the solutions.…”

“…Since RNA present in a given cell represents the portion of the genome that is expressed in that cell, northern analysis was considered a powerful tool for studying gene expression and measurement of a specific mRNA (Sabelli & Shewry, 1995). Nucleic acid hybridisation can be used to determine the concentration and the rate of synthesis of an individual RNA species in a complex situation (Williams, 1985). It is important to measure mRNA for two main reasons.…”