Нові генетичні конструкції KIN10-His/KIN11-His як інструмент для встановлення функціональної гомології протеїнкіназ SnRK1 та BSRK

О. Є. Краснопьорова, С. В. Ісаєнков, П. А. Карпов, А. І. Ємець

Анотація


Aim. The protein kinases SnRK1 from Arabidopsis thaliana are one of the key regulators of plant responses to different types of abiotic stresses. Many functions of these enzymes have not been studied yet. The possible functions of these protein kinases are regulation of cytoskeletal elements. To gain insight into molecular mechanisms of interaction of these enzymes with the cytoskeleton elements and discovery of potential substrates, the genetic constructs pGWB8-KIN10:His and pGWB8-KIN11:His for plant transformation were created. Methods. The coding sequences of KIN10 and KIN11 were cloned using gateway cloning system and other molecular-biological methods including PCR, RT-PCR. Results. The KIN10 and KIN11 His-tag fusions in genetic constructs pGWB8-KIN10:His and pGWB8-KIN11:His were created. Conclusions. We have created plasmid constructs pGWB8-KIN10:His аnd pGWB8-KIN11:His. According to bioinformatical analysis the KIN10 and KIN11 shared high level of homology with human BRSK1. Thus the KIN10 and KIN11might play important role in regulation of cytoskeleton. Created His-tag constructs can be used for identification of new substrates among cytoskeletal and other proteins.
Keywords: SnRK1, protein kinases, plasmid construct, сytoskeletal elements, polyhistidine-tag.


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