
Overview:
SPHK1 catalyzes the phosphorylation of sphingosine to form sphingosine-1-phosphate (SPP) which is a novel lipid messenger with both intracellular and extracellular functions. Various stimuli increase cellular levels of SPP by activation of SPHK1 (1). Competitive inhibitors of SPHK1 block formation of SPP and selectively inhibit cellular proliferation induced by a variety of factors. SPHK1/sphingosine-1-phosphate pathway contributes to colon carcinogenesis. SPHK1 plays a key role in the generation and release of proinflammatory mediators triggered by anaphylatoxins from human macrophages and also is involved in the process of immune cell motility (2).
Gene Aliases:
SPHK
Genbank Number:
BC008040
References:
1. Xia, P. et al: Sphingosine kinase interacts with TRAF2 and dissects tumor necrosis factor-alpha signaling. J. Biol. Chem. 2777996-8003, 2002. 2. Pappu, R. et al: Promotion of lymphocyte egress into blood and lymph by distinct sources of sphingosine-1-phosphate. Science 316295-298, 2007.
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实验非常不顺,想构建CDNA文库,但是从mRNA开始屡次失败,考虑主要是纯化过程中损失过多。想从总RNA入手,但是不知道实验步骤。不知那位大侠能提供总RNA建库的实验步骤。另外我现在手头有OligoDT,RT酶,苦于没有第二连合成试剂盒,不知道能否用普通PCR试剂盒Teq酶替代第二链合成过程中的DNA聚合酶I。好像有种方法合成第二链时,不需要另外的引物,利用降解的RNA作引物即可,我想直接设定PCR两个循环,合成第二链,不知方法可行否?愁啊,等着毕业,时间紧急,恳请帮忙。谢谢。

