牙鲆(Paralichthys olivaceus)糖原合成酶激酶-3 (GSK3)不同亚型的克隆及表达研究

The Cloning and Expression Analysis of Different Glycogen Synthase Kinase-3 (GSK3) Subtypes on Paralichthys olivaceus

糖原合成酶激酶-3 (Glycogen Synthase Kinase-3, GSK3)主要有GSK-3α和GSK-3β两种亚型,二者在催化活性区域具有高度的同源性。GSK3β可以参与包括Wnt信号在内的多个信号通路的调控,影响正常细胞的生存并参与肿瘤的发生与发展。本文以牙鲆为材料,通过基因克隆得到GSK-3α,GSK-3β和GSK3β-like 3个亚型,NCBI序列比对并经NJ法构建系统进化树,结果显示GSK3β-like亚型是海洋鱼类所特有的。同时,Megalign蛋白序列比对及SMART结构预测显示3个不同亚型的功能结构域高度同源;但qRT-PCR结果显示,GSK3β在胚胎发育时期的表达量明显高于另外两个亚型,这可能与GSK3β在多种生物学过程中发挥主要作用相关。

Glycogen synthase kinase-3 mainly includes two subtypes of GSK-3α and GSK-3β, whose catalytic activity area is highly homologous. GSK-3β can participate in multiple signaling pathways, including Wnt signal, affect the survival of normal cells and participate in the occurrence and development of the tumor. In this paper, we got GSK-3α, GSK-3β and GSK3β-like subtypes of Paralichthys olivaceus through gene cloning. NCBI Blast and NJ method built the system evolutionary tree;results showed that the GSK3β-like subtypes are peculiar to the marine fish. Meanwhile, Megalign protein sequence alignment and SMART structure prediction revealed that the three different subtypes are highly homologous in functional domain. But, the qRT-PCR results showed that GSK-3β presents significantly higher expression than the other two subtypes during embryonic development, which may be related to that GSK-3β plays a major role in a variety of biological processes.