
Overview:
Microtubule affinity-regulating kinase 3 (MARK3) is a member of the PAR-1/MARK kinase family which play critical roles in polarity and cell cycle control and are regulated by 14-3-3 scaffolding proteins, as well as the LKB1 tumour suppressor kinase and atypical protein kinase C (PKC)(1). MARK3 is a dual-specificity protein kinase that controls entry into mitosis by dephosphorylating CDC2. MARK3 seems to be a positive regulator of the beta-catenin pathway and an inhibitor of the JNK pathway. MARK3, a regulator of polarity, is also a modulator of Wnt-beta-catenin signalling, indicating a link between two important developmental pathways (2).
Gene Aliases:
KP78, CTAK1, PAR1A
Genbank Number:
NM_002376
References:
1. Goransson, O. et al: Regulation of the polarity kinases PAR-1/MARK by 14-3-3 interaction and Phosphorylation. J. Cell Sci. 2006;119(19):4059-70. 2. Kato, T. et al: Isolation of a novel human gene, MARKL1, homologous to MARK3 and its involvement in hepatocellular carcinogenesis. Neoplasia. 2001;3(1):4-9.
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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。

