Download e-book for iPad: Cell physiology: molecular dynamics by Henry Tedeschi

By Henry Tedeschi

ISBN-10: 0126851506

ISBN-13: 9780126851502

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Any of these can be targeted to specific intracellular compartments. GFP attached to repressor molecules has permitted identifying the position of operators in the study of gene expression in living cells (see Chapter 3). Like any other fluorescent probe, GFP and GFP mutants can be used in fluorescent recovery after photobleaching (FRAP) studies. One added feature is that the bleaching is reversible in GFP mutants with two excitation peaks. , 1997). , 2001, box 1) between different mutants of GFP (for other applications of FRET, see Chapter 4).

These procedures allow the quantitative evaluation of gene expression in real time, even with very small amounts of RNA. , 2000) can also be used to study protein-DNA interactions. How can you detect the binding of a protein to the DNA? A genetic approach to study the binding of proteins to DNA and RNA is discussed in Section IIB, an extension of the two-hybrid technique of detecting proteinprotein binding. This section will discuss biochemical aproaches. Apart from the older ultracentrifugation techniques that can detect a shift in molecular size by changes in sedimentation rates, two techniques have been predominantly used.

Binding properties The determination of the rate and binding of components, particularly proteins, has become increasingly important. g. , 1999). The role of protein-protein interactions are many. Enzymatic reactions have frequently been found to involve complexes, and hence binding of proteins to other proteins. For example, RNA polymerase II can function as a complex of 12 subunits. However, further studies found that it is comprised of 55 subunits to reconstitute the native transcriptional activity (see Koleske and Young, 1994).

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Cell physiology: molecular dynamics by Henry Tedeschi


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