
Overview:
PIP5K1B (phosphatidylinositol-4-phosphate 5-kinase, type I, beta) is a member of the phosphatidylinositol-4-phosphate 5-kinase family. PIP5K1B gene contains 17 exons and spans more than 300 kb. The seventeenth exon was found by RT-PCR, and are derived from the 3-prime untranslated region of the PRKACG gene which is located on 9q13 approximately 3 kb downstream of the STM7.I 3-prime untranslated region (1).The overexpression of PIP5K1B in COS-7 cells induces an increase in short actin fibers and a decrease in actin stress fibers(2).
Gene Aliases:
MSS4; STM7
Genbank Number:
NM_003558
References:
1. Pook, M. A.et.al: Exon-intron structure of a 2.7-kb transcript of the STM7 gene with phosphatidylinositol-4-phosphate 5-kinase activity. Genomics 42: 170-172, 1997.2. Ishihara, H.et.al: Type I phosphatidylinositol-4-phosphate 5-kinases: cloning of the third isoform and deletion/substitution analysis of members of this novel lipid kinase family. J. Biol. Chem. 273: 8741-8748, 1998.
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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。

