The Construction Of Eukaryotic Expression System And Primary Functional Research Of Human Neuritin

Abstract: Neurotrophic factors are growth factors or cytokines that are inducible polypeptides and permit intercellular communication. Neurotrophic factors can modulate the development of growth, maintenance of function ,and differention of neuronal population of nervous system, so they play an important role in supporting the injured nervous system and neurodegerative disorders. An explosion of information about neurotrophic factors is setting the stage for significant advances in neural disease therapy .Neuritin was a novel neurotrophic factor which was first reported by Nedivi as Candidate Plasticity-related genes15, on nature in 1993 ,then it was named by Israel scientist Naeve in 1997. Previous studies demonstrated that Neuritin can induce neurite growth, promote the outgrowth and arborization of neurite. Both BDNF and neuronal activity can active the expression of neuritin. It suggest that the neuritin may be a down-regulated co-factor of BDNF and neuronal activity . Neuritin applied foreground in nerve regenerative field, but the function and molecular mechanism of Neuritin is not very clearly so far.At present ,a human neuritin cDNA was cloned and the structure and potential function of its protein was predicted by bioinformatics . On the basis of past work, we aim to construct eukaryotic expression system and do the primary research of neuritin .Firstly, the coding sequence of human neuritin was cloned into pcDNA4.0 eukaryotic expression vector by using genetic recombination technique .After analysis by restriction enzyme digestion and DNA sequencing , the recombinant pcDNA4.0-neuritin plasmid was transfected into PC12 cells by using cationic lipid (Tfx-20). Finally, the expression of neuritin was determined by immunocytochemistry .The result showed that neuritin reading frame was correct. The recombinant plasmid can express at mRNA level and produce active Neuritin protein in eukaryotic cells .The research applied a foundmental basis for the function and mechanism study of neuritin in future.