Expression and localization of proteins in mammalian cells

Abstract

The transfer of genes and expression of recombinant proteins is one of the main methods for discovery in the quest for understanding as biology moves from genomes through to proteomes, and finally to working cells and organisms. The techniques and tools for the transfer and engineering of genetic material and for the detection and analysis of expressed proteins are evolving rapidly alongside the burgeoning databases they have to explore. Physical, chemical, and biologically based vectors increasingly enable gene transfer into mammalian cells for research and therapeutic purposes. The emergence of fluorescent protein tags now allows sophisticated tracking and functional analysis of expressed proteins. In this review, we summarize the methods and scientific tools contributing to this technology. Finally, the push is on to develop high‐throughput and computational methods for large‐scale protein expression, localization, and analysis, and here we set the stage by providing updates on the coming technology in these fields.

Visualizing and clustering high throughput sub-cellular localization imaging

“…The sequencing of numerous genomes and subsequent identification of the encoded proteome has created the need for large-scale systematic approaches to understand the functions of the tens of thousands of proteins at the cellular level [1,2]. High-throughput automated fluorescent microscope imaging technologies enable the experimental determination of a protein's sub-cellular localization and its dynamic trafficking within a range of cellular contexts.…”

Subcellular Localization of Mammalian Type II Membrane Proteins

“…This directed approach allows analysis of the function of each protein and yields an array of experimental data that are subject to experimental variations, such as the cell type analyzed, method of protein detection, and type and position of protein tag. This approach provides vital information about the specifics of individual proteins; however, it fails to address the global understanding of protein sorting as the extent of experimental variation prevents the direct comparison of results (7).…”