
Overview:
VLK or vertebrate lonesome kinase is a protein kinase which is first expressed in E-cadherin-positive anterior visceral endoderm and mesendoderm, and later its expression is confined to E-cadherin-negative mesenchyme. VLK regulates the rate of protein export from the Golgi and thereby plays an important role in the formation of functional stroma by mesenchymal cells (1). Targeted disruption of VLK leads to a defect in lung development and delayed ossification of endochondral bone. VLK deficient mice display neonatal lethality due to respiratory failure, with a suckling defect arising from a cleft palate. VLK is required for the appropriate timing of flat proliferative chondrocyte differentiation (2).
Gene Aliases:
SGK493; PKDCC
Genbank Number:
NM_138370
References:
1. Kinoshita. et.al: The novel protein kinase Vlk is essential for stromal function of mesenchymal cells.Development 136: 2069-2079, 2009. 2. Imuta, Y. et al: Short limbs, cleft palate, and delayed formation of flat proliferative chondrocytes in mice with targeted disruption of a putative protein kinase gene, Pkdcc (AW548124). Dev Dyn. 2009 Jan; 238(1):210-22.
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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。

