
Overview:
The PI3K comprises of a 110 kDa catalytic subunit and a 85 kDa regulatory subunit. A number of isoforms of the 110 kDa catalytic subunit and the 85 kDa regulatory subunit exist in cells. p85a modulates the interaction between PI3K and platelet-derived growth factor receptor (1). Furthermore, estrogen receptor isoform ER-alpha binds in a ligand-dependent manner to the p85-alpha regulatory subunit of PI3K. Stimulation with estrogen increases ER-alpha-associated PI3K activity, leading to the activation of protein kinase B/AKT and endothelial nitric oxide synthase (eNOS) (2).
Gene Aliases:
PI3K catalytic Domain a, PIK3R1, GRB1, p85-ALPHA
Genbank Number:
NM_181523
References:
1. Skolnik, E. Y. Et al: Cloning of PI3-kinase associated p85 utilizing a novel method for expression/cloning of target proteins for receptor tyrosine kinases. Cell 65: 83-90, 1991.2. Simoncini, T. et al: Interaction of oestrogen receptor with the regulatory subunit of phosphatidylinositol-3-OH kinase. Nature 407: 538-541, 2000.
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植物材料 用最精确的方法,称取不超过100mg的植物材料,置于处理过的研钵,加入液氮进行研磨 将研磨得到的粉末,快速转移至无RNase,并经过液氮冷却的2mL离心管(自备),
但是,这样做的前提的,RNA提得要好,浓度要高,这样免得RNA降解到不能用了。因为在有二价阳离子的(DNAseI buffer含有)情况下,RNA在60度以上时必然发生非酶促降解。
想搞清一个问题,如何区别是因为small RNA 引起的调控呢?还是tRNA引起的调控,有没有有效的分离这两种RNA的办法呢?
样品处理:
a. 植物组织:取新鲜或-70℃冻存100mg组织在液氮中研磨,把粉末加入到1ml裂解液中混匀。
b. 动物组织:取新鲜或-70℃冻存100mg组织加1ml裂解液,用组织研磨杵或匀浆器匀浆处理。
c. 贴壁细胞:直接在培养板中加入裂解液裂解细胞,每106细胞加1ml 裂解液。用取样器吹打混匀。

