Cloning And Prokaryotic Expression Of Potassium Channel Protein SKT1 From Patato

Potassium is one of the essential elements for plant growth and development,and is involved in many important physiological functions in plants.Potato is a kind of K-favoring crops,potassium nutrition can not only improve the photosynthetic efficiency of potato,but also affect the synthesis and transport of photosynthetic products,affecting the quality and yield of potato.Therefore,the accumulation of potassium in plants plays an important role in the growth and development of plants.The C-terminus of sktl gene containing 1758 bp was cloned by polymerase chain reaction.Then it was sub-cloned into pET30-TEV/LIC vector and was transformed into E.coli BL21(DE3)for expression of SKT1 recombinant protein.Unfortunately,SKT1 protein was existed in inclusion body.The inclusion body was isolated,dissolved with 6 mol/L guanidine hydrochloride,then refolded and purified by Nickel affinity chromatography.The purified target protein was identified by Western blot.More than 40%of inclusion body was refolded after dissolution and the SKT1 protein reached a purity of more than 90%after purification.In this study,the gene of potassium channel protein SKT1 was cloned from patota Solanum tuberosum L.Favorita,and SKT1 protein was expressed in E.coli,in order to study its crystal structure,function,and the interaction between SKT1 and other protein during potassium ion absorption process.