Cloning, tissue distribution, expression pattern, and function of porcine maternal embryonic leucine zipper kinase.

Background

Mrs. embryonic leucine zipper kinase (Melk) is an atypical member of snf1 / AMPK family of serine-threonine kinases, are involved in physiological and pathological processes vary, including cell proliferation, apoptosis, embryogenesis, cancer Mouse Recombinant Proteins treatment resistance, and RNA processing. Melk is highly expressed in human cancers and is associated with a more aggressive form of astrocytoma, glioblastoma, breast cancer, and melanoma until now, no information about swine Melk (pMELK) have been reported.

method

In this study, pMELK coding sequence was cloned from pig spleen and characterized. We also quantitatively determined the expression of Melk in 11 isolated from pig tissue and subcellular localization is determined as stated in the swine umbilical vein endothelial cells (SUVEC) as a fusion protein. In addition, we report the functional characterization of proteins pMELK regarding its role in apoptosis.

sequencing analysis showed that the full-length of pMELK was 2072 bp with 17 exons, encoding 655 amino acids, including S-TKC conserved domain. PMELK comparison with ten other mammal species from their orthologous sequences showed> 91% homology and evolutionary distance <0.05, indicating that the Melk highly conserved in evolution.

Melk relative quantification of expression in 11 tissue samples isolated from piglets 30-day-old showed expression in all organs tested Melk and its highest expression in the superficial inguinal lymph nodes. Build a plasmid named pEGFP-Melk and Melk GFP-fusion protein successfully expressed in SUVECs. fluorescence microscopy revealed the subcellular distribution of GFP-fusion protein Melk restricted to the cytoplasm.

Cloning, tissue distribution, expression pattern, and function of porcine maternal embryonic leucine zipper kinase

About function, Flow cytometry analysis showed that overexpression of GFP-pMELK in Improving cell SUVEC Staurosporine (STS) induced apoptosis, but did not differ significantly. The pMELK protein was also found to interact with swine BCL-G and temporary transfection of the recombinant plasmid pCMV-HA-pMELK be SUVEC cells Multi-species Recombinant Proteins stably express GFP-pBCL-inhibited G protein-G induced apoptosis pBCL significantly.The this study provided useful information about pMELK basic details and functions in apoptosis offer potential new molecular model for intervention by the disease and diseases associated with human Melk and BCL-G.

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