Isolation and Purification of Proteins

Authored by: Leland J. Cseke , Peter B. Kaufman , Ara Kirakosyan , Margaret V. Westfall

Handbook of Molecular and Cellular Methods in Biology and Medicine

Print publication date:  December  2011
Online publication date:  April  2016

Print ISBN: 9781420069389
eBook ISBN: 9781439881958
Adobe ISBN:

10.1201/b11351-19

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Abstract

Proteins and/or enzymes are the final products of gene expression. Although DNA stores the genetic information, it is the proteins that determine the shape and structure of a cell, a tissue, an organ, and the intact organism. Enzymes, which are usually proteins in nature, control the expression of genes and the development of an organism. Overall, proteins constitute half of the dry weight of the cell. In recent years, proteins and enzymes have become one of the core parts of molecular biology studies. Protein identification is important not only for detecting novel proteins, but also for monitoring protein expression in genetically modified animal models and/or after gene or cell transfer. A number of molecular biology studies start with the identification of a novel gene or protein(s) and then determine how the function or particular phenotype of an organism responds to changes in its expression. These types of experiments often utilize genetic screening of cDNA libraries followed by the cloning of a specific gene and then bacterial and eukaryotic expression systems to identify and characterize the associated protein(s) of interest. In addition, proteins/enzymes are involved in broad molecular biology studies such as the interaction between the cis-element and the trans-factor, DNA/protein interaction in gel retardation and footprinting, screening of cDNA libraries as probes, immunoblotting or precipitation, and in vitro translation. 1 6

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