In vivo functions of histone acetylation/deacetylation in Tup1p repression and Gcn5p activation.
In vivo functions of histone acetylation/deacetylation in Tup1p repression and Gcn5p activation.
复制标题
组蛋白乙酰化/脱乙酰化在 Tup1p 抑制和 Gcn5p 激活中的体内功能。
DOI：
10.1101/sqb.1998.63.459
复制
发表时间：
1998
期刊：
影响因子：
--
通讯作者：
Roth,SY
中科院分区：
文献类型：
--
作者：
Edmondson,DG;Zhang,W;Watson,A;Xu,W;Bone,JR;Yu,Y;Stillman,D;Roth,SY
方法：免疫斑点印迹法。将合成肽的系列稀释液在PVDF或硝酸纤维素膜上斑点印迹。然后用印度墨水对印迹进行染色，以验证已经加载或处理了等量的肽用于如前所述的远Western（Edmondson等人，1996）。简言之，将印迹在0.05%Tween-20的磷酸盐缓冲盐水（PBS）中封闭2小时，然后用1%牛血清白蛋白（BSA）的PBS溶液封闭2小时。用PBS短暂冲洗印迹，然后在含有Tup 1 p探针的PBS中的1%山羊血清、0.3% BSA中孵育。在[35 S]甲硫氨酸存在下，通过Tup 1 p的体外翻译产生探针。孵育2小时后，将印迹在PBS中洗涤4次，每次5分钟，使其干燥，然后暴露于X射线胶片或磷光成像仪屏幕。用ImageQuant™软件（Molecular Dynamics）进行定量。
METHODSFar-Western dot blots. Serial dilutions of synthetic peptides were dot-blotted on PVDF or nitrocellulose membranes. The blots were then stained with India ink to verify that equivalent amounts of peptides had been loaded or processed for far-Western as described previously (Edmondson et al. 1996). Briefly, blots were blocked for 2 hours in 0.05% Tween-20 in phosphatebuffered saline (PBS), followed by 1% bovine serum albumin (BSA) in PBS for 2 hours. The blots were briefly rinsed with PBS and then incubated in 1% goat serum, 0.3% BSA in PBS containing the Tup1p probe. The probe was generated by in vitro translation of Tup1p in the presence of [35S] methionine. After a 2-hour incubation, the blots were washed four times for 5 minutes each in PBS, allowed to dry, and then exposed to X-ray film or phosphoimager screens. Quantitation was performed with ImageQuant™ software (Molecular Dynamics).
登录
查看更多内容
体外酵母 α2 蛋白指导的转录抑制
影响因子：
64.8
作者：
B. M. Herschbach;M. Arnaud;A. Johnson
通讯作者：
A. Johnson
DOI：
10.1111/j.1432-1033.1989.tb14530.x
发表时间：
1989-01-15
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
TURNER, BM;FELLOWS, G
通讯作者：
FELLOWS, G
影响因子：
10.5
作者：
TZAMARIAS, D;STRUHL, K
通讯作者：
STRUHL, K
组蛋白 H4 的氨基末端突变会破坏稳定的核小体定位和酵母 α2 阻遏物的完全抑制。
影响因子：
10.5
作者：
S. Roth;M. Shimizu;L. Johnson;M. Grunstein;R. Simpson
通讯作者：
R. Simpson
DOI：
10.1074/jbc.272.17.11193
发表时间：
1997-04
期刊：
The Journal of Biological Chemistry
影响因子：
--
作者：
M. Redd;M. Arnaud;A. Johnson
通讯作者：
M. Redd;M. Arnaud;A. Johnson