
Overview:
UBE2D2(UBC4) Protein or ubiquitin-conjugating enzyme E2D 2 is a member of the E2 ubiquitin-conjugating enzyme family which functions in the ubiquitination of the tumor-suppressor protein p53, which is induced by an E3 ubiquitin-protein ligase. UBE2D2 specifically ubiquitinates E6AP and in vivo inhibition of UBE2D2 can lead to inhibition of E6-stimulated p53 degradation (1). UBE2D2 could conjugate ubiquitin to target proteins in an E6AP-dependent manner (2).
Gene Aliases:
UBC4; PUBC1; UBC4/5; UBCH5B; E2(17)KB2
Genbank Number:
NM_001093036
References:
1. Rolfe, M.et.al: Reconstitution of p53-ubiquitylation reactions from purified components: the role of human ubiquitin-conjugating enzyme UBC4 and E6-associated protein (E6AP). Proc. Nat. Acad. Sci. 92: 3264-3268, 1995. 2. Jensen, J. P.et.al: Identification of a family of closely related human ubiquitin conjugating enzymes. J. Biol. Chem. 270: 30408-30414, 1995.
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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。

