Tgf-beta Activated Kinase-like Gene Cloning And Preliminary Function Study

Transforming growth factor ?activated kinase (TAK1) may stimulate the stress-activated kinases, SAPK1 and SAPK2, leading to the activation of SAPK signaling transduction pathway, via phosphorylation of the MAP kinase kinases MKK3, MKK4 and MKK6, in cells stimulated by transforming growth factor β (TGF β). TAKl is also involved in the signal transduction pathways of interleukin 1 (IL1) and tumour necrosis factor (TNF). TAKl plays a crucial role in biological processes such as cell division, differention, development or extracellular matrix formation.By large-scale sequencing analysis of the human fetal brain cDNA library and bioinformatics analysis, we isolated a novel human TAK1 like (TAKL) gene and had some researches on the structure and function of TAKL. The TAKL cDNA was 1997 base pair in length, encoding a putative protein of 242 amino acids, which shared a homology with human, mouse, and Xenopus TAKl. The TAKL gene was located in human chromosome 21q21. Reverse transcription PCR revealed that TAKL gene was expressed ubiquitously in human adult and fetal tissues and expressed highly in breast carcinoma GI-101, colon adenocarcinoma GI-112, and prostatic adenocarcinoma PCS . Northern blot analysis revealed that the TAKL mRNA was expressed predominantly in peripheral blood leukocyte. The transcript 2.0 kb was consistent with the expected size. The constructed TAKL-pQE30 plasmid was expressed highly in E.coli with the induction of IPTG. The recombined protein was purified with Ni-NTA Superflow chromatography.