Titlebook: p53 Protocols; Sumitra Deb,Swati Palit Deb Book 2003 Humana Press 2003

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Book 2003this protein. Several unique properties of p53 posed a challenge to understa- ing its normal function in the initial phase of its research. The low levels of p53 in normal cells, its stabilization under situations of genotoxic stress, induction of growth arrest, and apoptosis with stabilization of t

evanescent

commonsense

Purification of Recombinant p53 from Sf9 Insect Cells,h with no flanking sequences and its expression is driven by the baculovirus polyhedron promoter. We also describe how to concentrate the p53 up to 0.9 mg/mL. By gel filtration analysis, we demonstrate that 20% of the p53 forms a tetramer, and 80% forms a monomer. In a DNA binding assay known as the

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Prologue

reperfusion

BURSA

,Real-Time Polymerase Chain Reaction Quantitation of Relative Expression of Genes Modulated by p53 Using SYBR® Green I,nalysis as a target of p53 transrepression and mutant p53 transactivation. A p53-null cell line derived from lung carcinoma, H1299, was infected with recombinant adenovirus expressing wild-type (WT) p53, mutant p53-D281G, or β-galactosidase as a control. After 24 h of infection, RNA was harvested an

FLAIL

Transactivation and Transrepression Studies with p53, and p73). Once a gene of interest is identified, its presumptive promoter region can be cloned upstream of a luciferase gene in a plasmid. The most common reason for transfection experiments is to study gene expression patterns in the presence or absence of a particular gene product (e.g., p53). Th

变化

Methods to Study p53-Repressed Promoters,n also function as a sequence-specific transcriptional repressor of a separate set of genes. However, elucidation of the mechanism whereby p53 functions as a transcriptional repressor has been obscured by the use of artificial assays to measure this activity; these assays include transient transfect