
Overview:
NEK 2 is closely related in its catalytic domain to the serine/threonine protein kinase NIMA of Aspergillus nidulans that is required for entry into mitosis and may function in parallel to the universal mitotic inducer p34cdc2. Like NIMA, the NEK2 protein is almost undetectable during G1 but accumulated progressively throughout S, reaching maximal levels in late G2 (1). NEK2 is shown to be expressed most abundantly in the testis of the adult tissues examined being localized to the nucleus (2).
Gene Aliases:
NLK1; HsPK21; NEK2A
Genbank Number:
NM_002497
References:
1. Schultz, SJ. et al: A. Cell cycle-dependent expression of Nek2, a novel human protein kinase related to the NIMA mitotic regulator of Aspergillus nidulans. Cell Growth Differ. 1994 Jun;5(6):625-35.2. Fry, AM. et al: Substrate specificity and cell cycle regulation of the Nek2 protein kinase, a potential human homolog of the mitotic regulator NIMA of Aspergillus nidulans. J Biol Chem. 1995 May 26; 270(21):12899-905.
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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。

