Cloning,Expression And Purification Of Novel Potential Protein Kinase-based Herbicide Targets

The herbicide plays an important role in the agricultural production because of its economic,fast and easily operation.However,as the long-term use of the herbicide with the single mechanism and target,the resistance problem and the environment problem as well as non-target organism's security problem of the weed become increasingly serious.So,to research more environmentally friendly,low toxicity,efficient herbicides have become an important topic nowadays.In this paper,we constructed 10 recombinant protein kinase plasmids based on the potential herbicide targets of the protein kinase gene which were screened from the genome sequence of the model plant arabidopsis thaliana,and purified 5 recombinant proteins by optimizing the expression and purification conditions.Finally,by high-performance liquid chromatography method,we detected the consumption of ATP and the generation of ADP in the process of the kinase reaction and confirmed the purified kinases have appropriate biological activity.The result laid a solid foundation for the future evaluation of these kinases structural and functional study.