DNA物理图谱的制备.PPT

酵母双杂交系统 Yeast Two-hybrid System(interaction trap) A System to Identify Proteins Involved in Specific Protein-protein Interactions Protein-protein interactions Protein-protein interactions are intrinsic to virtually every cellular process ranging from DNA replication, transcription, splicing and translation, to secretion, cell cycle control, intermediary metabolism, formation of cellular macrostructures and enzymatic complexes. The formation of large cellular structures such as the cytoskeleton, the nuclear scaffold, and the mitotic spindle result from complex interactions between proteins. Relatively smaller structures such as nuclear pores, centrosomes and kinetochores are beginning to be characterized and, in each case, protein-protein interactions seem to play a crucial role. 酵母单杂交系统Yeast one-hybrid system  酵母单杂交的原理与应用实例 The Mechanism and Application of Yeast One-hybrid System 生物工程进展 2001年04期陈峰 , 李洁 , 张贵友 , 刘强 许多诱导型基因的表达,都受特定的转录因子和顺式元件调控.要阐明各种信号传递途径与基因表达调控的机理,克隆和鉴定转录因子是关键.近年来酵母单杂交方法被广泛应用于克隆和鉴定各种动植物的转录因子,本文以拟南芥DREB转录因子的克隆为例,介绍酵母单杂交方法的原理和具体应用.  Transcriptional regulation of the NPT2 gene by dietary phosphate Kidney International (2001) 60, 412–415; Ken-Ichi?Miyamoto and Mikiko?Itho Transcriptional regulation of the NPT2 gene by dietary phosphate. Dietary phosphate (Pi) is an important regulator for renal Pi reabsorption. The type II sodium-dependent phosphate (Na/Pi) cotransporters (NPT2) are located at the apical membranes of renal proximal tubular cells and major functional transporters associated with renal Pi reabsorption. The yeast one-hybrid system was used to clone a transcription factor that binds to a specific sequence (Pi response element) in the promoter of the NPT2 gene. Two cDNA clones that encoded protein of the mouse transcription factor E3 (TFE3) were isolated. TFE3 may participate in the transcriptional regulation of the NPT2 gene by dietary Pi. Abbreviations: bp, base pair; EMSA, electrophoretic mobility shift assay; kb, kilo